Antiviral heterocyclic compounds

ABSTRACT

The present invention discloses compounds of Formula (I), or pharmaceutically acceptable salts, esters, or prodrugs thereof: 
     
       
         
         
             
             
         
       
     
     which inhibit Human Respiratory Syncytial Virus (HRSV) or Human Metapneumovirus (HMPV) inhibitors. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HRSV or HMPV infection. The invention also relates to methods of treating an HRSV or HMPV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

RELATED APPLICATIONS

This application is a continuation-in-part of U.S. application Ser. No.17/679,746, filed Feb. 24, 2022, which claims the benefit of U.S.Provisional Application No. 63/154,318, filed Feb. 26, 2021, U.S.Provisional Application No. 63/168,705, filed Mar. 31, 2021, U.S.Provisional Application No. 63/171,895, filed Apr. 7, 2021, and U.S.Provisional Application No. 63/293,339, filed Dec. 23, 2021. The entireteachings of the above applications are incorporated herein byreference.

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceuticalcompositions useful as Respiratory Syncytial Virus (RSV) inhibitors andHuman Metapneumovirus (HMPV) inhibitors.

BACKGROUND OF THE INVENTION

Human respiratory syncytial virus (HRSV) is a negative sense virus,containing a non-segmented, single-stranded linear RNA genome. As aParamyxovirus of two serotypes in the genus Pneumoviridae, HRSV contains10 genes that encode for 11 proteins. The nucleocapsid protein (N), theRNA polymerase protein (L), the phosphoprotein (P) and the transcriptionanti-termination factor (M2-1) along with the RNA genome make up theribonucleoprotein (RNP) complex. Several small-molecule compounds havebeen shown to target the RNP complex. Additionally, the fusion protein(F), paramount for viral attachment to the host, has been extensivelystudied. High resolution structures of the F protein interacting withinhibitors have been attained, while structural studies with the Nprotein are earlier in development. A direct result of the HRSV proteinstudies and research, the F protein, L protein and N protein have beenthe major focus of drug discovery efforts.

The increased effort in HRSV drug discovery is a result of HRSV beingthe leading cause of acute lower respiratory infections (ALRI) inpatients of all ages. In addition to respiratory infections, patientpopulations at high risk during HRSV infections include the elderly,immunocompromised, children up to the age of two and patients withchronic obstructive pulmonary disorder (COPD) or chronic heart failure(CHF). HRSV was found over four years to cause 177,500 hospitaladmissions and 14,000 deaths in the U.S. elderly population. It iswell-known that almost all children will be infected with HRSV in thefirst 3 years after birth and HRSV infection is more severe in prematureinfants. In fact, HRSV is the most common cause of bronchiolitis andpneumonia in infants under the age of one in the U.S. It is estimatedthat approximately 3.2 million hospitalizations and 66,000 deathsworldwide in children less than 5 years old are due to HRSV. HRSV hasbeen associated with more deaths of infants below one year old and moreinfant hospitalizations than influenza.

HRSV infection can also affect healthy individuals and repeated HRSVinfections even over the course of two months can occur. Symptoms aresimilar to colds in healthy individuals, however fever, wheezing, rapidand difficult breathing, and cyanosis occur in more severe cases.Currently, the treatment options for HRSV infection are quite limitedand there is no vaccine due to unsuccessful attempts to date.Palivizumab is a monoclonal antibody that is approved for prophylacticuse, but its use is limited due to its high price. Palivizumab isgenerally only used for high risk infants, such as premature infants orthose with cardiac/lung disease, but has been only 60% effective inreducing hospitalizations. Ribavirin is approved as an inhalationtreatment option, but its effectiveness is limited and there are safetyconcerns associated with it. Taking into account the treatment options,and the consistent seasonality of the HRSV epidemic, the development ofnew therapeutic agents for the treatment of HRSV is desirable.

There have been several RSV fusion inhibitors that have been disclosedin the following publications: WO2010/103306, WO2012/068622,WO2013/096681, WO2014/060411, WO2013/186995, WO2013/186334, WO2013/186332, WO 2012 080451, WO 2012/080450, WO2012/080449, WO2012/080447, WO 2012/080446, WO 2015/110446, WO 2017/009316, J. MedChem. 2015, 58, 1630-1643, Bioorg. Med Chem. Lett., 2015, 25, 976-981and Nat. Commun., 2017, 8, 167. Examples of other N-protein inhibitorsfor treatment of HRSV have been disclosed in the following publications:WO 2004/026843, J. Med Chem. 2006, 49, 2311-2319, and J. Med Chem. 2007,50, 1685-1692. Examples of L-protein inhibitors for HRSV have beendisclosed in the following publications: WO 2011/005842, WO 2005/042530,Antiviral Res. 2005, 65, 125-131, and Bioorg. Med. Chem. Lett. 2013, 23,6789-6793. Examples of nucleosides/polymerase inhibitors have beendisclosed in the following publications: WO 2011/005842, WO 2013/242525,WO 2014/031784, WO 2015/026792, WO 2016/0055791, WO 2016/138158 and J.Med. Chem. 2015, 58, 1862-1878.

Likewise, human metapneumovirus (HMPV), a negative-sense,single-stranded RNA enveloped virus, that belongs to the Pneumoviridaefamily and Metapneumovirus genus discovered by van Den Hoogen in 2001,is also a common cause of acute lower respiratory tract infections(ALRTIs). Although often mild, this virus can be serious andlife-threatening in high-risk groups, such as children under the age of5 years, elderly adults over the age of 65 years, and adults withunderlying disease (e.g., Chronic Obstructive Pulmonary Disease (COPD),asthma, congestive heart failure, or diabetes). In healthy adults overthe age of 65 years, the annual incidence rate of HMPV infection is1.2/1,000, and 38% of disease (e.g., COPD), and individuals are twice aslikely to have symptomatic disease and requirement for medical care. Inimmunocompromised individuals, HMPV is responsible for 6% of totalrespiratory infections in lung transplants and 3% of lower respiratoryinfections associated with stem cell transplant. HMPV infection is alsothought to be associated with acute graft rejection.

Like HRSV, infection is thought to attach to the target cell via theglycoprotein (G) protein interactions and followed by fusion via the Fprotein. HMPV L protein sequence is homologous to HRSV L protein.

HMPV infection is the second most common cause of lower respiratorytract infection in children (behind HRSV) and also problematic for theelderly population. There are 4 subtypes of HMPV found in clinicalisolates (A1, A2, B1 and B2). Reinfection occurs throughout childhoodfollowing initial infection. No therapeutics are currently available forHMPV infection.

Taking into account the seasonality and predictability of the HRSV andHMPV epidemics, HRSV epidemics in elderly institutions, and the severityof infection in high risk infants, the need for a potent and effectivetreatment for HRSV and HMPV is clear. The present invention hasidentified compounds that are heterocyclic molecules that are potentagainst HRSV-A/B and HMPV. The invention includes methods to preparethese molecules, methods for the RSV cell-based assay, the HMPV-GFPcell-based assay, the HMPV-TN/1501/A1 cell-based assay, andsmall-molecules that have potential to treat HRSV/HMPV infection.

SUMMARY OF THE INVENTION

The present invention provides compounds represented by Formula (I), andpharmaceutically acceptable salts, esters and prodrugs thereof that canbe used to treat or prevent viral (particularly HRSV or HMPV) infection:

wherein:A is selected from the group consisting of:

1) optionally substituted aryl; and

2) optionally substituted heteroaryl;

R₁ and R₂ are each independently selected from the group consisting of:

1) hydrogen;

2) fluorine; and

3) optionally substituted —C₁-C₆ alkyl;

alternatively, R₁ and R₂ are taken together with the carbon atom towhich they are attached to form an optionally substituted 3- to6-membered ring;Z is selected from the group consisting of:

1) hydrogen;

2) halogen; and

3) optionally substituted —C₁-C₆ alkyl;

W is selected from the group consisting of:

1) hydrogen;

2) optionally substituted —C₁-C₆ alkoxy;

3) optionally substituted —C₁-C₆ alkyl; and

4) optionally substituted —C₃-C₆ cycloalkyl;

G is selected from the group consisting of:

1) —C(O)OR₁₂;

2) —C(O)NR₁₁R₁₂;

3) optionally substituted —C₁-C₆ alkyl-CN;

4) optionally substituted —C₁-C₆ alkyl-C(O)NR₁₁R₁₂;

5) optionally substituted —C₁-C₆ alkyl-C(O)NR₁₁S(O)₂R₁₂;

6) optionally substituted —C₁-C₆ alkyl-OC(O)NR₁₁R₁₂;

7) optionally substituted —C₁-C₆ alkyl-NHR₁₃;

8) optionally substituted —C₁-C₆ alkyl-NHC(O)R₁₃;

n is 1, 2 or 3; preferably n is 1 or 2;Y is O, S(O)₂, or NR₁₄;E is selected from the group consisting of:

1) optionally substituted aryl;

2) optionally substituted heteroaryl;

3) optionally substituted 3- to 8-membered heterocyclic, and

4) optionally substituted alkynyl;

R₃ is hydroxy or fluorine;R₄ is selected from the group consisting of:

1) hydrogen;

2) optionally substituted —C₁-C₆ alkyl;

3) optionally substituted —C₃-C₈ cycloalkyl; and

4) optionally substituted 3- to 8-membered heterocyclic;

R₁₁ at each occurrence is independently selected from the groupconsisting of:

1) hydrogen;

2) optionally substituted —C₁-C₈-alkyl;

3) optionally substituted —C₃-C₈-cycloalkyl;

4) optionally substituted 4- to 8-membered heterocyclic;

5) optionally substituted aryl;

6) optionally substituted arylalkyl;

7) optionally substituted heteroaryl; and

8) optionally substituted heteroarylalkyl;

R₁₂ at each occurrence is independently selected from the groupconsisting of:

1) hydrogen;

2) optionally substituted —C₁-C₈-alkyl;

3) optionally substituted —C₃-C₈-cycloalkyl;

4) optionally substituted 4- to 8-membered heterocyclic;

5) optionally substituted aryl;

6) optionally substituted arylalkyl;

7) optionally substituted heteroaryl; and

8) optionally substituted heteroarylalkyl;

alternatively, R₁₁ and R₁₂ are taken together with the nitrogen atom towhich they are attached to form a 3- to 12-membered heterocyclic ring,preferably the said 3- to 12-membered heterocyclic ring is, but notlimited to morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, and,azetidine;R₁₃ at each occurrence is independently selected from the groupconsisting of:

1) Optionally substituted —C₁-C₈ alkyl;

2) Optionally substituted —C₃-C₈ cycloalkyl;

3) Optionally substituted 4- to 8-membered heterocyclic;

4) Optionally substituted aryl;

5) Optionally substituted arylalkyl;

6) Optionally substituted heteroaryl; and

7) Optionally substituted heteroarylalkyl; and

R₁₄ is selected from:

1) hydrogen;

2) optionally substituted —C₁-C₈-alkyl; and

3) optionally substituted —C₃-C₈-cycloalkyl;

Each preferred group stated above can be taken in combination with one,any or all other preferred groups.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment of the present invention is a compound of Formula (I)as described above, or a pharmaceutically acceptable salt thereof.

In certain embodiments of the compounds of Formula (I), Y is O.

In certain embodiments of the compounds of Formula (I), Y is O, and n is1 or 2.

In certain embodiments of the compounds of Formula (I), R₁ is hydrogenor F.

In certain embodiments of the compounds of Formula (I), R₂ is hydrogenor F.

In certain embodiments of the compounds of Formula (I), Z is hydrogen,Cl or F.

In certain embodiments of the compounds of Formula (I), R₁ is hydrogen,R₂ is hydrogen, and Z is hydrogen.

In certain embodiments of the compounds of Formula (I), W is optionallysubstituted methyl, optionally substituted ethyl, or optionallysubstituted cyclopropyl.

In certain embodiments of the compounds of Formula (I), W iscyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂, or —CF₃.

In certain embodiments of the compounds of Formula (I), R₃ is —OH.

In certain embodiments of the compounds of Formula (I), R₄ is optionallysubstituted methyl or optionally substituted cyclopropyl.

In certain embodiments of the compounds of Formula (I), R₃ is OH, and R₄is CF₃ or cyclopropyl.

In certain embodiments of the compounds of Formula (I), R₁ is hydrogen,R₂ is hydrogen, R₃ is OH, and R₄ is CF₃.

In certain embodiments of the compounds of Formula (I), Gis-C(O)NR₁₁R₁₂. In certain embodiments, G is —C(O)NH₂.

In certain embodiments of the compounds of Formula (I), G is —CH₂NHR₁₃,—CH₂C(O)NR₁₁R₁₂, —CH₂NHC(O)R₁₃, —CH₂OC(O)NR₁₁R₁₂, —CH₂CN, or—CH₂C(O)NR₁₁S(O)₂R₁₂. In certain embodiments, G is —CH₂C(O)NH₂.

In certain embodiments of the compounds of Formula (I), A is selectedfrom one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted.

In certain embodiments of the compounds of Formula (I), A is selectedfrom the groups set forth below,

wherein each of these groups is optionally substituted.

In certain embodiments of the compounds of Formula (I), A is selectedfrom the groups set forth below,

In certain embodiments of the compounds of Formula (I), A is

wherein Ra is hydrogen, halogen, —CN, —NO₂, —OR₁₁, —NR₁₁R₁₂,—NR₁₁C(O)R₁₂, —NR₁₁S(O)₂R₁₂, —S(O)₂R₁₂, —S(O)₂NR₁₁R₁₂, —NR₁₁C(O)NR₁₁R₁₂,—C(O)R₁₁, —C(O)OR₁₁, —C(O)NR₁₁R₁₂, optionally substituted —C₁-C₆ alkyl,optionally substituted —C₃-C₈-cycloalkyl, optionally substituted 3- to8-membered heterocyclic, optionally substituted aryl, or optionallysubstituted heteroaryl; Rb and Rb′ are each independently selected fromhydrogen, halogen, —OR₁₁, —NR₁₁R₁₂, optionally substituted —C₁-C₆-alkyl,optionally substituted —C₃-C₈-cycloalkyl, optionally substituted 3- to8-membered heterocyclic, optionally substituted aryl, and optionallysubstituted heteroaryl. Alternatively, Rb and Rb′ are taken togetherwith the carbon atoms to which they are attached to form a 4- to7-membered ring fused with the phenyl ring.

In certain embodiments of the compounds of Formula (I), E is optionallysubstituted aryl, preferably optionally substituted phenyl.

In certain embodiments of the compounds of Formula (I), E is selectedfrom one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted.

In certain embodiments of the compounds of Formula (I), E is selectedfrom the groups set forth below,

In certain embodiments of the compounds of Formula (I), E is selectedfrom the groups set forth below,

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (Ia) or Formula (Ib), or a pharmaceuticallyacceptable salt, ester or prodrug thereof:

wherein A, R₁, R₂, Z, W, G, n, Y, E, R₃, and R₄ are as previouslydefined.

In a preferred embodiment, the compound of Formula (I) has thestereochemistry shown in Formula (Ib).

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (II), or a pharmaceutically acceptable salt,ester, or prodrug thereof:

wherein A, R₁, R₂, W, G, n, E, R₃, and R₄ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (III) or a pharmaceutically acceptable salt,ester, or prodrug thereof:

wherein A, W, G, n, Y, E, R₃, and R₄ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (IV-1)˜(IV-2), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein A, W, G, Y, E, R₃, and R₄ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (V-1)˜(V-4), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein A, W, G, E, R₁₄, R₃, and R₄ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (VI-1)˜(VI-4), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein A, W, G, E, R₁₄, R₃, and R₄ are as previously defined.Preferably, W is optionally substituted methyl; more preferably, W is—CH₃ or —CF₃.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (VII-1)˜(VII-12), or apharmaceutically acceptable salt, ester, or prodrug thereof:

wherein A, W, E, R₁₁, R₁₂, R₁₃, R₁₄, R₃, and R₄ are as previouslydefined. Preferably, W is cyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂, or—CF₃; more preferably, W is —CH₃ or —CF₃.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (VIII-1)˜(VIII-12), or apharmaceutically acceptable salt, ester, or prodrug thereof:

wherein A, W, E, R₁₁, R₁₂, R₁₃, R₁₄, R₃, and R₄ are as previouslydefined. Preferably, W is cyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂, or—CF₃; more preferably, W is —CH₃ or —CF₃.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (IX-1)˜(IX-4), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein each R₂₁ is independently optionally substituted methyl, halo,—CN, —OR₁₁, or —NR₁₁R₁₂; m is 0, 1, 2, 3, 4 or 5; A, W, G, Ru, R₁₂, R₁₄,R₃, and R₄ are as previously defined. Preferably, each R₂₁ isindependently halo or optionally substituted methyl, and m is 1 or 2.More preferably, each R₂₁ is independently —F, —Cl, —CN, —CF₃, —CH₂F or—CHF₂, and m is 1 or 2.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (X-1)˜(X-4), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein R₂₁, m, A, W, G, R₁₄, R₃, and R₄ are as previously defined.Preferably, each R₂₁ is independently halo or optionally substitutedmethyl, and m is 1 or 2.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XII-1)˜(XII-12), or apharmaceutically acceptable salt, ester, or prodrug thereof:

wherein m′ is 0, 1, or 2. A, R₃, R₄, R₂₁, R₁₁, R₁₂ and R₁₃ are aspreviously defined. Preferably m′ is 2.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XIII-1)˜(XIII-12), or apharmaceutically acceptable salt, ester, or prodrug thereof:

wherein R₂₁, m′, A, R₃, R₄, Rn, R₁₂, and R₁₃ are as previously defined.Preferably m′ is 2.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XIV-1)˜(XIV-6), or a pharmaceuticallyacceptable salt, ester or prodrug thereof:

wherein each R₂₂ is independently halo; —CN; —NO₂, —OR₁₁; —NR₁₁R₁₂;—NR₁₁C(O)R₁₂; —NR₁₁S(O)₂R₁₂; —S(O)₂R₁₂; —S(O)₂NR₁₁R₁₂, —NR₁₁C(O)NR₁₁R₁₂;—C(O)R₁₁, —C(O)OR₁₁; —C(O)NR₁₁R₁₂; optionally substituted —C₁-C₆ alkyl;optionally substituted —C₃-C₈-cycloalkyl; optionally substituted 3- to8-membered heterocyclic; optionally substituted aryl; or optionallysubstituted heteroaryl, and W, m′, R₃, R₄, R₂₁, R₁, and R₁₂ are aspreviously defined. Preferably, R₂₁ is halogen, R₃ is —OH, R₄ is —CH₃,—CF₃, or cyclopropyl, and W is cyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂,or —CF₃. In certain embodiments, two adjacent R₂₂ groups are takentogether with the carbon atoms to which they are attached to form a 4-to 12-membered carbocyclic or heterocyclic, and which said 4- to12-membered carbocyclic or heterocyclic is fused with the phenyl orquinolinyl.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XV-1)˜(XV-6), or a pharmaceuticallyacceptable salt, ester or prodrug thereof:

wherein W, m′, R₃, R₄, R₂₁, R₂₂, R₁₁, and R₁₂ are as previously defined.Preferably, R₂₁ is halogen, R₃ is —OH, R₄ is —CH₃, —CF₃, or cyclopropyl,and W is cyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂, or —CF₃. In certainembodiment, two adjacent R₂₂ groups are taken together with the carbonatoms to which they are attached to form a 4- to 12-membered carbocyclicor heterocyclic ring, and which said 4- to 12-membered carbocyclic orheterocyclic is fused with the phenyl or quinolinyl.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XVI-1)˜(XVI-12), or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₂₃ is hydrogen; halo; —CN; —NO₂, —OR₁₁; —NR₁₁R₁₂; —NR₁₁C(O)R₁₂;—NR₁₁S(O)₂R₁₂; —S(O)₂R₁₂; —S(O)₂NR₁₁R₁₂, —NR₁₁C(O)NR₁₁R₁₂; —C(O)R₁₁,—C(O)OR₁₁; —C(O)NR₁₁R₁₂; optionally substituted —C₁-C₆ alkyl; optionallysubstituted —C₃-C₈-cycloalkyl; optionally substituted 3- to 8-memberedheterocyclic; optionally substituted aryl; or optionally substitutedheteroaryl; and W, R₂₁, R₂₂, m′, R₃, R₄, R₁₁, and R₁₂ are as previouslydefined. Preferably, R₂₁ is halogen, R₃ is —OH, R₄ is —CH₃, —CF₃, orcyclopropyl, and W is cyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂, or —CF₃.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XVII-1)˜(XVII-12), or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein W, R₂₁, R₂₂, R₂₃, m′, R₃, R₄, R₁₁, and R₁₂ are as previouslydefined. Preferably, R₂₁ is halogen, R₃ is —OH, R₄ is —CH₃, —CF₃, orcyclopropyl, and W is cyclopropyl, ethyl, —CH₃, —CH₂F, —CHF₂, or —CF₃.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XVIII-1)˜(XVIII-14), or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₁₄, R₂₁, and R₂₂ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XIX-1)˜(XIX-14), or apharmaceutically acceptable salt, ester, or prodrug thereof:

wherein R₁₄, R₂₁, and R₂₂ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XX-1)˜(XX-20), or a pharmaceuticallyacceptable salt, ester or prodrug thereof:

wherein each R₃₁ is independently optionally substituted —C₁-C₃ alkyl orhalo; R₃₂ is independently halo, —OR₁₁, —NR₁₁R₁₂, —NR₁₁C(O)R₁₂,—C(O)NR₁₁R₁₂, —C(O)R₁₁, optionally substituted —C₁-C₆ alkyl, oroptionally substituted —C₃-C₈-cycloalkyl; and R₁₁, R₁₂, and R₁₄ are aspreviously defined. Preferably, R₃₁ is halo, R₃₂ is halo, —NH₂,optionally substituted methyl, optionally substituted cyclopropyl, andR₁₄ is optionally substituted cyclopropyl.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XXI-1)˜(XXI-20), or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₃₁, R₃₂, R₁₁, R₁₂, and R₁₄ is as previously defined.Preferably, R₃₁ is halo, R₃₂ is halo, —NH₂, optionally substitutedmethyl, optionally substituted cyclopropyl, and R₁₄ is optionallysubstituted cyclopropyl.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XXII-1)˜(XXII-14), or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₂₄ is hydrogen or R₂₂; and W, R₄, R₁₄, R₂₁, and R₂₂ are aspreviously defined. Preferably, R₄ is optionally substituted —C₃-C₆cycloalkyl. More preferably, R₄ is optionally substituted cyclopropyl oroptionally substituted cyclobutyl.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formulae (XXIII-1)˜(XXIII-14), or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein W, R₄, R₁₄, R₂₁, R₂₂, and R₂₄ are as previously defined.Preferably, R₄ is optionally substituted —C₃-C₆ cycloalkyl. Morepreferably, R₄ is optionally substituted cyclopropyl or optionallysubstituted cyclobutyl, and W is optionally substituted methyl.

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (XXIV), or a pharmaceutically acceptable salt,ester, or prodrug thereof:

wherein A, E, R₃, and R₄ are each as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (XXIV), or a pharmaceutically acceptable salt,ester, or prodrug thereof, wherein R₃ is —OH; R₄ is optionallysubstituted methyl or optionally substituted —C₃-C₆ cycloalkyl; A isselected from:

and E is selected from:

or E is selected from

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (XXV), or a pharmaceutically acceptable salt,ester, or prodrug thereof:

wherein each R₄₁, R₄₂, R₄₃, R₄₄, and R₄₅ are each independently selectedform hydrogen, halogen, optionally substituted methyl, optionallysubstituted methoxyl or —CN. R₃, R₄, R₁₄, and R₂₄ are as previouslydefined. Preferably, R₃ is —OH; R₄ is optionally substituted methyl oroptionally substituted —C₃-C₆ cycloalkyl; R₁₄ is optionally substituted—C₃-C₆ cycloalkyl or optionally substituted methyl; and R₂₄ isoptionally substituted methoxyl or halogen.

In one embodiment of the present invention, the compound of Formula(XXV), or a pharmaceutically acceptable salt, ester or prodrug thereof,wherein R₄₂ and R₄₃ are taken together with the carbon atoms to whichthey are attached to form a fused 4- to 7-membered carbocyclic ring orheterocyclic ring.

In one embodiment of the present invention, the compound of Formula(XXV), or a pharmaceutically acceptable salt, ester or prodrug thereof,wherein R₄₁ and R₄₂ are taken together with the carbon atoms to whichthey are attached to form a fused 4- to 7-membered carbocyclic ring orheterocyclic ring.

In one embodiment of the present invention, the compound of Formula (I)is represented by one Formulae (XXVI-1)˜(XXVI-3), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein R₃, R₁₄, R₂₄, R₄₁, R₄₂, R₄₃, R₄₄, and R₄₅ are as previouslydefined.

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (XXVII), or a pharmaceutically acceptablesalt, ester, or prodrug thereof:

wherein R₁₄, R₂₄, R₄₁, R₄₂, R₄₃, R₄₄, and R₄₅ are as previously defined.

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (XXVIII), or a pharmaceutically acceptablesalt, ester, or prodrug thereof.

wherein R₁₄, R₂₄, R₄₁, R₄₂, R₄₃, R₄₄, and R₄₅ are as previously defined.

In one embodiment of the present invention, the compound of Formula(XXVII), or Formula (XXVIII), or a pharmaceutically acceptable salt,ester or prodrug thereof, wherein R₄₂ and R₄₃ are taken together withthe carbon atoms to which they are attached to form a fused 4- to7-membered carbocyclic ring or heterocyclic ring.

In one embodiment of the present invention, the compound of Formula(XXVII), or Formula (XXVIII), or a pharmaceutically acceptable salt,ester or prodrug thereof, wherein R₄₃ and R₄₄ are taken together withthe carbon atoms to which they are attached to form a fused 4- to7-membered carbocyclic ring or heterocyclic ring.

In one embodiment of the present invention, the compound of Formula (I)is represented by one of Formula (XXIV), Formula (XXV), Formulae(XXVI-1)˜(XXVI-3), Formula (XXVII), or Formula (XXVIII), or apharmaceutically acceptable salt, ester or prodrug, wherein R₁₄ isoptionally substituted —C₃-C₆ cycloalkyl or optionally substitutedmethyl; R₂₄ is optionally substituted methoxyl or halogen; and each R₄₁,R₄₂, R₄₃, R₄₄, and R₄₅ are each independently selected from hydrogen,—F, —Cl, —CH₃, —CF₃, —OCH₃, or —OCF₃.

In one embodiment of the present invention, the compound of Formula (I)is represented by Formula (XXIV), Formula (XXV), Formulae(XXVI-1)˜(XXVI-3), Formula (XXVII), or Formula (XXVIII), or apharmaceutically acceptable salt, ester or prodrug, wherein R₁₄ is—CHF₂,

R₂₄ is —F, —C, or —OCH₃; and

is selected from:

is selected from

In one embodiment of the present invention, the compound of Formula (I)is represented by Formulae (XXIX-1)˜(XXIX-3), or a pharmaceuticallyacceptable salt, ester, or prodrug thereof:

wherein R₄₆ is substituted methyl or optionally substituted cyclopropyl;R₄₇ is hydrogen, Cl, F, or optionally substituted methoxyl, R₄₁, R₄₂,R₄₃, R₄₄, and R₄₅ are as previously defined. In certain embodiments ofthe compounds of Formulas (XXIX-1), (XXIX-2) and (XXIX-3), R₄₆ iscyclopropyl, 1-fluorocyclopropyl or difluoromethyl.

In one embodiment of the present invention, the compound of one ofFormulae (XXIX-1)˜(XXIX-3), or a pharmaceutically acceptable salt, esteror prodrug thereof, wherein R₄₂ and R₄₃ are taken together with thecarbon atoms to which they are attached to form a fused 4- to 7-memberedcarbocyclic ring or heterocyclic ring.

In one embodiment of the present invention, the compound of one ofFormulae (XXIX-1) (XXIX-3), or a pharmaceutically acceptable salt, esteror prodrug thereof, wherein R₄₃ and R₄₄ are taken together with thecarbon atoms to which they are attached to form a fused 4- to 7-memberedcarbocyclic ring or heterocyclic ring.

It will be appreciated that the description of the present inventionherein should be construed in congruity with the laws and principles ofchemical bonding. In some instances, it may be necessary to remove ahydrogen atom in order to accommodate a substituent at any givenlocation.

It is intended that the definition of any substituent or variable (e.g.,R₁, R₂, etc.) at a particular location in a molecule be independent ofits definitions elsewhere in that molecule.

It will be yet appreciated that the compounds of the present inventionmay contain one or more asymmetric carbon atoms and may exist inracemic, diastereoisomeric, and optically active forms. It will still beappreciated that certain compounds of the present invention may exist indifferent tautomeric forms. All tautomers are contemplated to be withinthe scope of the present invention.

In certain embodiments, the present invention provides a method for theprevention or treatment of RSV activities and for treating RSV infectionin a subject in need thereof. The method comprises administering to thesubject a therapeutically effective amount of a compound of formula (I).

The present invention also provides the use of a compound of formula (I)for the preparation of a medicament for the prevention or treatment ofRSV.

Thus, in one embodiment, a compound of formula (I), or pharmaceuticallyacceptable salt thereof, is combined with a steroid anti-inflammatorycompound, for example budesonide or fluticasone. In a preferredembodiment, the steroid is administered in low doses to minimizeimmuno-suppressant effects. In another embodiment a compound of formula(I), or a pharmaceutically acceptable salt thereof, is combined with anon-steroid anti-inflammatory compound, for example leukotrieneantagonists such as Singulair (Merck) or Accolate (Astra Zeneca),phosphodiesterase 4 inhibitors such as roflumilast (Altana), TNF alphainhibitors such as Enbrel (Amgen), Remicade (Centocor), Humira (Abbott)or CDP870 (Celltech) or NSAIDS. In a further embodiment, a compound offormula (I) is combined with interleukin 8 or interleukin 9 inhibitors.The present invention thus also relates to a product containing acompound of formula (I), or a pharmaceutically acceptable salt thereof,and an anti-inflammatory compound for simultaneous, separate orsequential use in the treatment of RSV.

The present invention also relates to a combination of a compound offormula (I), or a pharmaceutically acceptable salt thereof, with ananti-influenza compound and the use of such a combination in thetreatment of concomitant RSV and influenza infections. The presentinvention thus also relates to a product containing a compound offormula (I), or a pharmaceutically acceptable salt thereof, and ananti-influenza compound for simultaneous, separate or sequential use inthe treatment of concomitant RSV and influenza infections. The compoundsof the invention may be administered in a variety of dosage forms. Thus,they can be administered orally, for example as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules.The compounds of the invention may also be administered parenterally,whether subcutaneously, intravenously, intramuscularly, intrasternally,transdermally or by infusion techniques. The compounds may also beadministered as suppositories.

In an embodiment, the compounds of the invention are administered byintranasal or intrabronchial administration. The present invention alsoprovides an inhaler or nebulizer containing a medicament which comprises(a) a derivative of the formula (I), as defined above, or apharmaceutically acceptable salt thereof, and (b) a pharmaceuticallyacceptable carrier or diluent.

The present invention also provides a pharmaceutical compositioncontaining such a benzodiazepine derivative, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier ordiluent.

The compounds of the invention are typically formulated foradministration with a pharmaceutically acceptable carrier or diluent.For example, solid oral forms may contain, together with the activecompound, diluents, e.g., lactose, dextrose, saccharose, cellulose, cornstarch or potato starch; lubricants, e.g., silica, talc, stearic acid,magnesium or calcium stearate, and/or polyethylene glycols; bindingagents; e.g., starches, arabic gums, gelatin, methylcellulose,carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents,e.g., starch, alginic acid, alginates or sodium starch glycolate;effervescing mixtures; dyestuffs; sweeteners; wetting agents, such aslecithin, polysorbates, laurylsulphates; and, in general, nontoxic andpharmacologically inactive substances used in pharmaceuticalformulations. Such pharmaceutical preparations may be manufactured inknown manner, for example, by means of mixing, granulating, tableting,sugar coating, or film coating processes.

Liquid dispersions for oral administration may be syrups, emulsions andsuspensions. The syrups may contain as carriers, for example, saccharoseor saccharose with glycerine and/or mannitol and/or sorbitol.

Suspensions and emulsions may contain as carrier, for example a naturalgum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol. The suspension orsolutions for intramuscular injections may contain, together with theactive compound, a pharmaceutically acceptable carrier, e.g., sterilewater, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and ifdesired, a suitable amount of lidocaine hydrochloride.

Solutions for injection or infusion may contain as carrier, for example,sterile water or preferably they may be in the form of sterile, aqueous,isotonic saline solutions.

The present invention also relates to the novel compounds, as definedabove; or a pharmaceutically acceptable salt thereof, for use in amethod of treating the human or animal body. The present invention alsorelates to a pharmaceutical composition comprising a novel compound asdefined above and a pharmaceutically acceptable diluant or carrier.Preferably, the pharmaceutical composition comprises a pharmaceuticallyacceptable salt of a novel compound as defined above. A pharmaceuticallyacceptable salt is as defined above. The novel compounds of theinvention are typically administered in the manner defined above and thecompounds are typically formulated for administration in the mannerdefined above.

Preferably, the pharmaceutical compositions comprise optically activeisomers of the novel compounds of the invention. Thus, for example,preferred novel compounds of the invention containing only one chiralcenter include an R enantiomer in substantially pure form, an Senantiomer in substantially pure form and enantiomeric mixtures whichcontain an excess of the R enantiomer or an excess of the S enantiomer.It is particularly preferred that pharmaceutical contains a compound ofthe invention which is a substantially pure optical isomer. For theavoidance of doubt, the novel compounds of the invention can, ifdesired, be used in the form of solvates.

Yet a further aspect of the present invention is a process of making anyof the compounds delineated herein employing any of the synthetic meansdelineated herein.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “aryl,” as used herein, refers to a mono-, bi-, or polycycliccarbocyclic ring system comprising at least one aromatic ring,including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl,indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system thatcomprises at least one aromatic ring. Polycyclic aryls can comprisefused rings, covalently attached rings or a combination thereof.

The term “heteroaryl,” as used herein, refers to a mono-, bi-, orpolycyclic aromatic radical having one or more ring atom selected fromS, O and N; and the remaining ring atoms are carbon, wherein any N or Scontained within the ring may be optionally oxidized. Heteroarylincludes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl,pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl,thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. A polycyclicheteroaryl can comprise fused rings, covalently attached rings or acombination thereof.

In accordance with the invention, aromatic groups can be substituted orunsubstituted.

The term “bicyclic aryl” or “bicyclic heteroaryl” refers to a ringsystem consisting of two rings wherein at least one ring is aromatic;and the two rings can be fused or covalently attached.

The term “alkyl” as used herein, refers to saturated, straight- orbranched-chain hydrocarbon radicals. “C₁-C₃ alkyl,” “C₁-C₆ alkyl,”“C₁-C₁₀ alkyl”C₂-C₄ alkyl,” or “C₃-C₆ alkyl,” refer to alkyl groupscontaining from one to three, one to six, one to ten carbon atoms, 2 to4 and 3 to 6 carbon atoms respectively. Examples of C₁-C₈ alkyl radicalsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl,n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl and octyl radicals.

The term “alkenyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon double bond bythe removal of a single hydrogen atom. “C₂-C₁₀ alkenyl,” “C₂-C₈alkenyl,” “C₂-C₄ alkenyl,” or “C₃-C₆ alkenyl,” refer to alkenyl groupscontaining from two to ten, two to eight, two to four or three to sixcarbon atoms respectively. Alkenyl groups include, but are not limitedto, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,heptenyl, octenyl, and the like.

The term “alkynyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon triple bond bythe removal of a single hydrogen atom. “C₂-C₁₀ alkynyl,” “C₂-C₈alkynyl,” “C₂-C₄ alkynyl,” or “C₃-C₆ alkynyl,” refer to alkynyl groupscontaining from two to ten, two to eight, two to four or three to sixcarbon atoms respectively. Representative alkynyl groups include, butare not limited to, for example, ethynyl, 1-propynyl, 1-butynyl,heptynyl, octynyl, and the like.

The term “cycloalkyl”, as used herein, refers to a monocyclic orpolycyclic saturated carbocyclic ring or a bi- or tri-cyclic groupfused, bridged or spiro system, and the carbon atoms may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic,iminic or oximic double bond. Preferred cycloalkyl groups include C₃-C₁₂cycloalkyl, C₃-C₆ cycloalkyl, C₃-C₈ cycloalkyl and C₄-C₇ cycloalkyl.Examples of C₃-C₁₂ cycloalkyl include, but not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl,4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl,spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, andthe like.

The term “cycloalkenyl”, as used herein, refers to monocyclic orpolycyclic carbocyclic ring or a bi- or tri-cyclic group fused, bridgedor spiro system having at least one carbon-carbon double bond and thecarbon atoms may be optionally oxo-substituted or optionally substitutedwith exocyclic olefinic, iminic or oximic double bond. Preferredcycloalkenyl groups include C₃-C₁₂ cycloalkenyl, C₃-C₈ cycloalkenyl orC₅-C₇ cycloalkenyl groups. Examples of C₃-C₁₂ cycloalkenyl include, butnot limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl,bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl,bicyclo[4.2.1]non-3-en-9-yl, and the like.

As used herein, the term “arylalkyl” means a functional group wherein analkylene chain is attached to an aryl group, e.g., —CH₂CH₂-phenyl. Theterm “substituted arylalkyl” means an arylalkyl functional group inwhich the aryl group is substituted. Similarly, the term“heteroarylalkyl” means a functional group wherein an alkylene chain isattached to a heteroaryl group. The term “substituted heteroarylalkyl”means a heteroarylalkyl functional group in which the heteroaryl groupis substituted.

As used herein, the term “alkoxy” employed alone or in combination withother terms means, unless otherwise stated, an alkyl group having thedesignated number of carbon atoms connected to the rest of the moleculevia an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy,2-propoxy (isopropoxy) and the higher homologs and isomers. Preferredalkoxy are (C₁-C₃) alkoxy.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclic and cycloalkenyl moiety described herein can also be analiphatic group or an alicyclic group.

An “aliphatic” group is a non-aromatic moiety comprised of anycombination of carbon atoms, hydrogen atoms, halogen atoms, oxygen,nitrogen or other atoms, and optionally contains one or more units ofunsaturation, e.g., double and/or triple bonds. Examples of aliphaticgroups are functional groups, such as alkyl, alkenyl, alkynyl, O, OH,NH, NH₂, C(O), S(O)₂, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH₂, S(O)₂NH,S(O)₂NH₂, NHC(O)NH₂, NHC(O)C(O)NH, NHS(O)₂NH, NHS(O)₂NH₂, C(O)NHS(O)₂,C(O)NHS(O)₂NH or C(O)NHS(O)₂NH₂, and the like, groups comprising one ormore functional groups, non-aromatic hydrocarbons (optionallysubstituted), and groups wherein one or more carbons of a non-aromatichydrocarbon (optionally substituted) is replaced by a functional group.Carbon atoms of an aliphatic group can be optionally oxo-substituted. Analiphatic group may be straight chained, branched, cyclic, or acombination thereof and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, as used herein, aliphaticgroups expressly include, for example, alkoxyalkyls, polyalkoxyalkyls,such as polyalkylene glycols, polyamines, and polyimines, for example.Aliphatic groups may be optionally substituted.

The term “carbocycle” or “carbocyclic” refers to a saturated, partiallyunsaturated or aromatic cyclic group in which each atom within the ringis carbon. Examples of cabocyclics include cycloalkyl, cycloalkenyl andaryl groups.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi- ortri-cyclic group fused, bridged or spiro system, where (i) each ringsystem contains at least one heteroatom independently selected fromoxygen, sulfur and nitrogen, (ii) each ring system can be saturated orunsaturated (iii) the nitrogen and sulfur heteroatoms may optionally beoxidized, (iv) the nitrogen heteroatom may optionally be quaternized,(v) any of the above rings may be fused to an aromatic ring, and (vi)the remaining ring atoms are carbon atoms which may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic,iminic or oximic double bond. Representative heterocycloalkyl groupsinclude, but are not limited to, 1,3-dioxolane, pyrrolidinyl,pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl,isothiazolidinyl, quinoxalinyl, pyridazinonyl,2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl,5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl,and tetrahydrofuryl. Such heterocyclic groups may be furthersubstituted. Heteroaryl or heterocyclic groups can be C-attached orN-attached (where possible).

It is understood that any alkyl, alkenyl, alkynyl, alicyclic,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphaticmoiety or the like, described herein can also be a divalent ormultivalent group when used as a linkage to connect two or more groupsor substituents, which can be at the same or different atom(s). One ofskill in the art can readily determine the valence of any such groupfrom the context in which it occurs.

The term “substituted” refers to substitution by independent replacementof one, two, or three or more of the hydrogen atoms with substituentsincluding, but not limited to, —F, —Cl, —Br, —I, —OH, —C₁-C₁₂-alkyl;—C₂-C₁₂-alkenyl, —C₂-C₁₂-alkynyl, —C₃-C₁₂-cycloalkyl, protected hydroxy,—NO₂, —N₃, —CN, —NH₂, protected amino, oxo, thioxo, —NH—C₁-C₁₂-alkyl,—NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl, —NH—C₃-C₁₂-cycloalkyl, —NH-aryl,—NH-heteroaryl, —NH-heterocycloalkyl, -dialkylamino, -diarylamino,-diheteroarylamino, —O—C₁-C₁₂-alkyl, —O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl,—O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl, —O-heterocycloalkyl,—C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl,—C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl, —C(O)— heteroaryl,—C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl,—CONH—C₂-C₈-alkynyl, —CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl,—CONH-heteroaryl, —CONH— heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl,—OCO₂—C₂-C₈-alkenyl, —OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl,—OCO₂-aryl, —OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —CO₂—C₁-C₁₂ alkyl,—CO₂—C₂-C₈ alkenyl, —CO₂—C₂-C₈ alkynyl, CO₂—C₃-C₁₂-cycloalkyl,—CO₂-aryl, CO₂-heteroaryl, CO₂-heterocyloalkyl, —OCONH₂,—OCONH—C₁-C₁₂-alkyl, —OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl,—OCONH—C₃-C₁₂-cycloalkyl, —OCONH-aryl, —OCONH-heteroaryl,—OCONH-heterocyclo-alkyl, —NHC(O)H, —NHC(O)—C₁-C₁₂-alkyl,—NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl, —NHC(O)—C₃-C₁₂-cycloalkyl,—NHC(O)-aryl, —NHC(O)-heteroaryl, —NHC(O)-heterocyclo-alkyl,—NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl, —NHCO₂— C₂-C₈-alkynyl,—NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl, —NHCO₂-heteroaryl,—NHCO₂-heterocycloalkyl, —NHC(O)NH₂, —NHC(O)NH—C₁-C₁₂-alkyl,—NHC(O)NH—C₂-C₈-alkenyl, —NHC(O)NH—C₂-C₈-alkynyl,—NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl, —NHC(O)NH-heteroaryl,—NHC(O)NH-heterocycloalkyl, NHC(S)NH₂, —NHC(S)NH—C₁-C₁₂-alkyl,—NHC(S)NH—C₂-C₈-alkenyl, —NHC(S)NH—C₂-C₈-alkynyl,—NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl, —NHC(S)NH-heteroaryl,—NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂, —NHC(NH)NH—C₁-C₁₂-alkyl,—NHC(NH)NH—C₂-C₈-alkenyl, —NHC(NH)NH—C₂-C₈-alkynyl,—NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl,—NHC(NH)NH-heterocycloalkyl, —NHC(NH)—C₁-C₁₂-alkyl,—NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl,—S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl, —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₈-alkenyl, —SO₂NH— C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthio-methyl. In certain embodiments, the substituents areindependently selected from halo, preferably C₁ and F; C₁-C₄-alkyl,preferably methyl and ethyl; halo-C₁-C₄-alkyl, such as fluoromethyl,difluoromethyl, and trifluoromethyl; C₂-C₄-alkenyl; halo-C₂-C₄-alkenyl;C₃-C₆-cycloalkyl, such as cyclopropyl; C₁-C₄-alkoxy, such as methoxy andethoxy; halo-C₁-C₄-alkoxy, such as fluoromethoxy, difluoromethoxy, andtrifluoromethoxy, —CN; —OH; NH₂; C₁-C₄-alkylamino; di(C₁-C₄-alkyl)amino;and NO₂. It is understood that the aryls, heteroaryls, alkyls, and thelike can be further substituted. In some cases, each substituent in asubstituted moiety is additionally optionally substituted when possiblewith one or more groups, each group being independently selected fromC₁-C₄-alkyl; —CF₃, —OCH₃, —OCF₃, —F, —Cl, —Br, —I, —OH, —NO₂, —CN, and—NH₂.

In certain embodiments, a substituted alkyl, alkenyl or alkoxy group issubstituted with one or more halogen atoms, preferably fluorine orchlorine atoms. Such substituted alkyl groups include fluoromethyl,difluoromethyl and trifluoromethyl. Such substituted alkoxy groupsinclude fluoromethoxy, difluoromethoxy and trifluoromethoxy.

The term “halo” or halogen” alone or as part of another substituent, asused herein, refers to a fluorine, chlorine, bromine, or iodine atom.

The term “optionally substituted”, as used herein, means that thereferenced group may be substituted or unsubstituted. In one embodiment,the referenced group is optionally substituted with zero substituents,i.e., the referenced group is unsubstituted. In another embodiment, thereferenced group is optionally substituted with one or more additionalgroup(s) individually and independently selected from groups describedherein.

The term “hydrogen” includes hydrogen and deuterium. In addition, therecitation of an atom includes other isotopes of that atom so long asthe resulting compound is pharmaceutically acceptable.

In certain embodiments, the compounds of each formula herein are definedto include isotopically labelled compounds. An “isotopically labelledcompound” is a compound in which at least one atomic position isenriched in a specific isotope of the designated element to a levelwhich is significantly greater than the natural abundance of thatisotope. For example, one or more hydrogen atom positions in a compoundcan be enriched with deuterium to a level which is significantly greaterthan the natural abundance of deuterium, for example, enrichment to alevel of at least 1%, preferably at least 20% or at least 50%. Such adeuterated compound may, for example, be metabolized more slowly thanits non-deuterated analog, and therefore exhibit a longer half-life whenadministered to a subject. Such compounds can synthesize using methodsknown in the art, for example by employing deuterated startingmaterials. Unless stated to the contrary, isotopically labelledcompounds are pharmaceutically acceptable.

The term “hydroxy activating group,” as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxyl groupso that it will depart during synthetic procedures such as in asubstitution or an elimination reaction. Examples of hydroxyl activatinggroup include, but are not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxyl,” as used herein, refers to a hydroxy groupactivated with a hydroxyl activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxyl protecting groups includebenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxy-carbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl,chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl,methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, allyl,benzyl, triphenyl-methyl (trityl), methoxymethyl, methylthiomethyl,benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl,trimethylsilyl, triisopropylsilyl, and the like.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “hydroxy prodrug group,” as used herein, refers to a promoietygroup which is known in the art to change the physicochemical, and hencethe biological properties of a parent drug in a transient manner bycovering or masking the hydroxy group. After said syntheticprocedure(s), the hydroxy prodrug group as described herein must becapable of reverting back to hydroxy group in vivo. Hydroxy prodruggroups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and thePharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York(1992) and in “Prodrugs of Alcohols and Phenols” by S. S. Dhareshwar andV. J. Stella, in Prodrugs Challenges and Rewards Part-2, (Biotechnology:Pharmaceutical Aspects), edited by V. J. Stella, et al, Springer andAAPSPress, 2007, pp 31-99.

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, methoxycarbonyl, t-butoxycarbonyl, 9-fluorenyl-methoxycarbonyl,benzyloxycarbonyl, and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such compounds are well known to those skilledin the art, and it will be obvious to those skilled in the art thatindividual solvents or mixtures thereof may be preferred for specificcompounds and reaction conditions, depending upon such factors as thesolubility of reagents, reactivity of reagents and preferred temperatureranges, for example. Further discussions of aprotic solvents may befound in organic chemistry textbooks or in specialized monographs, forexample: Organic Solvents Physical Properties and Methods ofPurification, 4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, N Y, 1986.

The term “protic solvent,” as used herein, refers to a solvent thattends to provide protons, such as an alcohol, for example, methanol,ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Suchsolvents are well known to those skilled in the art, and it will beobvious to those skilled in the art that individual solvents or mixturesthereof may be preferred for specific compounds and reaction conditions,depending upon such factors as the solubility of reagents, reactivity ofreagents and preferred temperature ranges, for example. Furtherdiscussions of protogenic solvents may be found in organic chemistrytextbooks or in specialized monographs, for example: Organic SolventsPhysical Properties and Methods of Purification, 4th ed., edited by JohnA. Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable,” as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the Formula herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley andSons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The term “subject,” as used herein, refers to an animal. Preferably, theanimal is a mammal. More preferably, the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. Tautomers may be incyclic or acyclic. The configuration of any carbon-carbon double bondappearing herein is selected for convenience only and is not intended todesignate a particular configuration unless the text so states; thus acarbon-carbon double bond or carbon-heteroatom double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

Certain compounds of the present invention may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present invention includes each conformationalisomer of these compounds and mixtures thereof.

As used herein, the term “pharmaceutically acceptable salt,” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentane-propionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

Pharmaceutically acceptable salts can also be prepared by deprotonationof the parent compound with a suitable base, thereby forming the anionicconjugate base of the parent compound. In such salts the counter ion isa cation. Suitable cations include ammonium and metal cations, such asalkali metal cations, including Li⁺, Na⁺, K⁺ and Cs⁺, and alkaline earthmetal cations, such as Mg²⁺ and Ca²⁺.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, esters ofC₁-C₆-alkanoic acids, such as acetate, propionate, butyrate and pivalateesters.

In certain embodiments, the invention provides pharmaceuticallyacceptable prodrugs of the compounds disclosed herein. The term“pharmaceutically acceptable prodrugs” as used herein refers to thoseprodrugs of the compounds formed by the process of the present inventionwhich are, within the scope of sound medical judgment, suitable for usein contact with the tissues of humans and lower animals with unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use,as well as the zwitterionic forms, where possible, of the compounds ofthe present invention. “Prodrug”, as used herein means a compound, whichis convertible in vivo by metabolic means (e.g. by hydrolysis) to affordany compound delineated by the formulae of the instant invention.Various forms of prodrugs are known in the art, for example, asdiscussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985);Widder, et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press(1985); Krogsgaard-Larsen, et al., (ed.). “Design and Application ofProdrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et al., Journal of Drug Deliver Reviews,8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq.(1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug DeliverySystems, American Chemical Society (1975); and Bernard Testa & JoachimMayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, ethyl succinate, phosphate esters,dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlinedin Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs ofhydroxy and amino groups are also included, as are carbonate prodrugs,sulfonate esters and sulfate esters of hydroxy groups. Derivatization ofhydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers wherein theacyl group may be an alkyl ester, optionally substituted with groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities, or where the acyl group is an amino acid ester asdescribed above, are also encompassed. Prodrugs of this type aredescribed in J. Med. Chem. 1996, 39, 10. Free amines can also bederivatized as amides, sulfonamides or phosphonamides. All of theseprodrug moieties may incorporate groups including but not limited toether, amine and carboxylic acid functionalities. In certainembodiments, a compound of the invention can incorporate two or moregroups that are metabolically removed in vivo to yield the active parentcompound.

The term “treating”, as used herein, means relieving, lessening,reducing, eliminating, modulating, or ameliorating, i.e., causingregression of the disease state or condition. Treating can also includeinhibiting, i.e., arresting the development, of an existing diseasestate or condition, and relieving or ameliorating, i.e., causingregression of an existing disease state or condition, for example whenthe disease state or condition may already be present.

The term “preventing”, as used herein means, to completely or almostcompletely stop a disease state or condition, from occurring in apatient or subject, especially when the patient or subject ispredisposed to such or at risk of contracting a disease state orcondition.

Additionally, the compounds of the present invention, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules. Nonlimitingexamples of hydrates include monohydrates, dihydrates, etc. Nonlimitingexamples of solvates include ethanol solvates, acetone solvates, etc.

“Solvates” means solvent addition forms that contain eitherstoichiometric or nonstoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent iswater, the solvate formed is a hydrate, when the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one of the substances in whichthe water retains its molecular state as H₂O, such combination beingable to form one or more hydrate.

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analog is acompound that is similar to or comparable in function and appearance tothe reference compound.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. Additionally, thevarious synthetic steps may be performed in an alternate sequence ororder to give the desired compounds. In addition, the solvents,temperatures, reaction durations, etc. delineated herein are forpurposes of illustration only and variation of the reaction conditionscan produce the desired bridged macrocyclic products of the presentinvention. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein include, for example, those described in R.Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d.Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser andFieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); andL. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, JohnWiley and Sons (1995).

The compounds of this invention may be modified by appending variousfunctionalities via synthetic means delineated herein to enhanceselective biological properties. Such modifications include those whichincrease biological penetration into a given biological system (e.g.,blood, lymphatic system, central nervous system), increase oralavailability, increase solubility to allow administration by injection,alter metabolism and alter rate of excretion.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose, any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one with ordinary skill inthe art. All publications, patents, published patent applications, andother references mentioned herein are hereby incorporated by referencein their entirety.

Abbreviations

Abbreviations which have been used in the descriptions of the schemesand the examples that follow are:

ACN for acetonitrile;AD-mix-β for(9S)-(9″S)-9,9″-[1,4-Phthalazinediylbis(oxy)]bis[10,11-dihydro-6′-methoxycinchonan];Bn for benzyl;BOP for (Benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate;BzCl for benzoyl chloride;mCPBA for meta-chloroperbenzoic acid;Cbz for benzyloxycarbonyl;CDI for carbonyldiimidazole;DAST for diethylaminosulfur trifluoride;

DBU for 1,8-Diazabicycloundec-7-ene;

DCE for dichloroethane;DCM for dichloromethane;Dess-Martin periodinane for1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one;DIAD for diisopropyl azodicarboxylate;DIBAL-H for diisobutylaluminum hydride;

DMAP for N,N-dimethylaminopyridine;

DME for 1,2-dimethoxyethane;DMF for N,N-dimethyl formamide;DMSO for dimethylsulfoxide;DPPA for diphenylphosphoryl azide or diphenyl phosphorylazidate;dppf for 1,1′-Bis(diphenylphosphino)ferrocene;EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimidehydrochloride;EA or EtOAc for ethyl acetate;Ghosez's reagent for 1-Chloro-N,N,2-trimethyl-1-propenylamine;HATU for O (7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate;HCl for hydrochloric acid;Hunig's base for diisopropylethylamine;PyBOP for (Benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate;LDA for Lithium diisopropylamine;Pd—C for palladium carbon;PE for petroleum ether;Ph for phenyl;RT for reverse transcription;RT-PCR for reverse transcription-polymerase chain reaction;TBME for tert-butyl methyl ether;TEA for triethylamine;Tf₂O for trifluoromethanesulfonic anhydride;TFA for trifluoroacetic acid;THE for tetrahydrofuran;(TMS)₂NH for hexamethyldisilazane;TBS for tert-Butyldimethylsilyl;TBDPS for tert-Butyldiphenylsilyl;TMS for trimethylsilyl;TPAP tetrapropylammonium perruthenate;TPP or PPh₃ for triphenylphosphine;Ts or tosyl for p-CH₃C₆H₄SO₂—;tBOC or Boc for tert-butyloxy carbonyl; andXantphos for 4,5-Bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene.

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared, which are intended as an illustration only and not to limitthe scope of the invention. Various changes and modifications to thedisclosed embodiments will be apparent to those skilled in the art andsuch changes and modifications including, without limitation, thoserelating to the chemical structures, substituents, derivatives, and/ormethods of the invention may be made without departing from the spiritof the invention and the scope of the appended claims.

Scheme 1 illustrates methods to prepare a compound of formula 11 fromcompounds 1 and 2, wherein n=1, 2 or 3; P is hydroxy protecting group;Ar is E; and E is as previously defined. Alkylation of the hydroxypyridine 1 with hydroxy epoxide using Mitsunobu reaction conditionsaffords epoxide 4. Alternatively, hydroxy epoxide is converted to 3which has a leaving group such as but not limited to, tosyl andmethanlsulfonyl followed by alkylation in the presence of base such asbut not limited to, K₂CO₃ and Cs₂CO₃, provides 4. Intramolecular epoxideopening mediated by base such as but not limited to, LDA, producescompound 5. Hydroxy group compound 5 is protected with proper protectinggroup such as but not limited to, TBDPS and TBS, affords compound 6.Trifluomethyl ketone 7 is obtained from iodine-magnesium exchange ofcompound 6 followed by addition of ester such as but not limited to,ethyl 2,2,2-trifluoroacetate. Trifluoromethyl ketone 7 in cross-coupledwith various metal coupling partners 8, but not limited to, boronicacids, boronic esters, organotin reagents, organozinc reagents,organomagnesium reagents, organo silicon reagents or the like catalyzedby appropriate Pd, Ni, Cu or the like catalyst to afford compound 9.Nitromethane addition in the presence of base such as but not limitedto, K₂CO₃ and Cs₂CO₃, to compound 9 affords compound 10. Reduction ofnitro group with reducing reagents such as but not limited to, zinc andacetic acid, produces key intermediate 11.

As seen in scheme 2, wherein Ar₁ is A; Ar is E; R is R₁₁; n is 1, 2 or3; and A, E, R₁₁ are as previously defined. Key intermediate 11 iscoupled with various carboxylic acids to afford amide 14. Amide 14 isthen reacted with a variety of electrophiles to produce various ethers,esters and carbamates of formula 15. Amide 14 is also oxidized to analdehyde 16, and then reductive amination provides a variety of amines17. The hydroxyl of —CH₂OH in amide 14 is converted to cyanomethyl 18via activation followed by cyanation. Compound 14-1 is furthertransformed to acetamide 19 in the presence of a catalyst such as, butnot limited to, Parkin's catalyst.

As seen in scheme 3, wherein Ar₁ is A; Ar is E; R is R₁₁; and A, E, R₁₁are as previously defined. An aldehyde 16 is converted to the benzylprotected amine through reductive amination. Hydrogenolysis affords thefree amine 20. Lastly, displacement with a variety of electrophilesgives N-substituted compounds 21.

As seen in Scheme 4, wherein Ar₁ is A; Ar is E; R′ is —C₁-C₆ alkyl,—C₃-C₆ cycloalkyl, aryl, or heteroaryl; n is 1, 2 or 3; and A and E areas previously defined. After oxidation of aldehyde 16 to acid 22, whichis further converted to amides 23 and sulfonamides 24 using commonmethods such as but not limited to, HATU and DIPEA. From therediversification to a variety of esters and amides is conducted.

Scheme 5 illustrates another method to prepare a compound of formula 11,wherein Ar is E; P is a hydroxy protecting group; n is 1, 2 or 3; and Eis as previously defined. Ketone 9 is converted to compound of formula26 via olefination. Alternatively, 26 is obtained from; 1) 6 viacross-coupling with metal coupling partner 6-1, which can be, but is notlimited to, a boronic acid, a boronic ester, an organotin reagent, anorganozinc reagent, an organomagnesium reagent, an organosilicon reagentor the like catalyzed by appropriate Pd, Ni, Cu or the like catalyst toafford compound 25; 2) compound 25 is converted to compound 26 aspreviously described method in scheme 1. With 26 in hand, Compounds offormula 27 are prepared by dihydroxylation followed by epoxideformation. Epoxide opening of compound 27 with amine equivalent such asbut not limited to, NH₄OH and NH₃, provides compounds of formula 11.

Scheme 6 illustrates another method to prepare a compound of formula 23,wherein Ar₁ is A; Ar is E; R′ is —C₁-C₆ alkyl, —C₃-C₆ cycloalkyl, aryl,or heteroaryl; n is 1, 2 or 3; and A and E are as previously defined.Amine 11 is protected with a protecting group such as but not limitedto, Boc and Cbz. After deprotection of hydroxy protecting group,subsequent oxidations provide acid 30. Compound 30 is coupled withvarious amines to provide amide 31. Deprotection of amine protectinggroup followed by subsequent amide formation affords compounds offormula 23.

Scheme 7 illustrates an additional route for the synthesis of thedesired compounds. The difference for this route is that it starts withoxidation and amide coupling to install the amide 33 at the beginning ofthe synthesis. Sequential vinylation and arylation afford thebis-coupled product 34. Asymmetric dihydroxylation followed byactivation and substitution affords the amino alcohol precursor. Lastly,amide coupling with the respective aryl acid produces the desiredcompounds depicted by compound 36.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art, and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Certain synthetic procedures useful in the preparation of compounds ofthe invention are disclosed in U.S. patent application Ser. No.16/930,622, the contents of which are incorporated by reference hereinin their entirety.

Example 1

Example 1 Step a

A solution of 3-bromo-4-hydroxybenzoate (16 g, 69.25 mmol), Cs₂CO₃ (68g, 207.75 mmol), KI (46 g, 277.00 mmol) and bromocyclopropane (21 g,173.12 mmol) in NMP (30 mL) was stirred for 16 hours at 180° C. in aParr reactor. The resulting solution was diluted with water andextracted with EtOAC. The combined organics were dried and concentrated.The resulting solution was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to afford desired product as a yellow solid(3 g, 22%). ESI-MS m/z: 257.05 [M+H]⁺. (Methyl ester product was alsoisolated and used).

Example 1 Step b

A solution of the compound from step a (250 mg, 0.98 mmol), Pd(dppf)Cl₂(142 mg, 0.19 mmol),2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (425mg, 1.94 mmol), H₂O (0.1 mL) and Cs₂CO₃ (950 mg, 2.91 mmol) in dioxane(3 mL) was stirred for 2 hours at 90° C. under N₂ atmosphere. Theresulting solution was purified by reverse phase C18 columnchromatography (MeOH/0.1% FA in H₂O) to afford the desired product as awhite solid (180 mg, 68%). ESI-MS m/z: 270.15 [M+H]⁺.

Example 1 Step c

To a 2-dram vial equipped with a stir bar was added amine (30 mg, 0.075mmol), acid (19.18 mg, 0.075 mmol), and the material was dissolved inDMF (0.2 M). Hunig's base (0.053 mL, 0.30 mmol) was added and the vialwas cooled to 0° C. HATU (43 mg, 0.113 mmol) was added, the reactionstirred for 10 minutes, warmed to room temperature and monitored by LCMS(1 hr). The reaction was diluted with EtOAc and quenched with water. Theaqueous was extracted with EtOAc and DCM/MeOH with a phase separatorcartridge and concentrated. The material was purified by prep-HPLC20-90%, MeCN/Water, 25 min to afford the title compound as a white solid(23.6 mg, 48%). ESI-MS m/z: 651.25 [M+H]⁺.

Example 2

Example 2 Step a

To a 50 mL round-bottom flask equipped with a stir bar was added methyl8-hydroxyquinoline-6-carboxylate (1.500 g, 7.38 mmol) and potassiumcarbonate (2.040 g, 14.76 mmol), and the solids were dissolved in DMF(0.5 M). tert-Butyl 2-bromoacetate (1.308 ml, 8.86 mmol) was then added,the reaction stirred at 40° C. and monitored by LCMS (2 hr). Thereaction was cooled to r.t., diluted with EtOAc and quenched with water.Aqueous was extracted with EtOAc, and the combined organics dried,filtered, and concentrated. The residue was purified by automated columnchromatography (silica gel, R_(f)=0.27 in 50% ethyl acetate in hexanes)to afford a white, solid (1.93 g, 82%). ESI-MS m/z: 262.0 [M+H]⁺.

Example 2 Step b

To a 100 mL round-bottom flask containing step a (1.93 g, 6.08 mmol) wasadded a stir bar, and the solid dissolved in DCM (0.5 M). The flask wascooled to 0° C., and TFA (4.69 ml, 60.8 mmol) was added. The reactionwas stirred for 10 minutes, warmed to room temperature and monitored byLCMS (added 5.0 eq. more TFA after 3 hr, 5.5 hr total). The mixture wasquenched with water and diluted with DCM. Solid precipitates. Furtherdiluted with DCM and stirred vigorously for 10 minutes. The solid wascollected by filtration and washed multiple times with DCM and driedunder high vacuum to afford a light brown, fluffy solid (2.21 g, 97%).ESI-MS m/z: 262.0 [M+H]⁺.

Example 2 Step c

To a 40 mL vial equipped with a stir bar was added step b (125 mg, 0.333mmol). The solid was dissolved in DMF and cooled to 0° C. DIPEA (407 μl,2.332 mmol) was added followed by 1-methylcyclopropan-1-aminehydrochloride (124 mg, 1.148 mmol). PyBOP (260 mg, 0.500 mmol) was thenadded in one portion, the reaction stirred for 10 minutes, warmed toroom temperature and monitored by LCMS (1.5 hr). The reaction dilutedwas with EtOAc and quenched with water. The aqueous layer was extractedwith EtOAc, with a phase separator cartridge, and the combined organicswere concentrated. The residue was purified by automated columnchromatography (silica gel, 0-20% methanol in dichloromethane) the titlecompound (98 mg, 82%). ESI-MS m/z: 216.0 [M+H]⁺.

Example 2 Step d

General hydrolysis notes: In some cases, the reaction was heated to 45°C. to force material into solution and accelerate hydrolysis. Afterhydrolysis, the product was isolated by precipitation. If noprecipitate, the product was either extracted, or the aqueous solutionwas concentrated (material dried and used crude). The major MS' m/z forall these compounds is C—C cleavage: ESI-MS m/z: 202.0 [M+H]⁺.

To a 20 mL vial containing Example 2 Step c (9 mg, 0.312 mmol) was addeda stir bar. The compound was dissolved in MeOH, THE and Water (0.2 M,2:1:1). Lithium hydroxide hydrate (62 mg, 1.56 mmol) was added, thereaction stirred at room temperature and monitored by LCMS. The stir barwas removed, and the vial cooled to 0° C. The reaction was acidifiedwith 2 M HCl, and the pH brought to around 4-5 (used 1M NaOH if tooacidic). The product was extracted 3× with 10% MeOH/DCM with a phaseseparator and concentrated. Dried on high vacuum to afford the titlecompound (45 mg, 50%). ESI-MS m/z: 202.0 [M+H]⁺.

Example 2 Step e

The following example was prepared using the same procedures as Example1 Step c, with the corresponding acid from step d and amine HCl salt (25mg) coupling partners. The residue was purified by Gilson prep-HPLC(20-90%, MeCN/Water, 25 min) to afford the title compound (8 mg, 20%)ESI-MS m/z: 682.2.

Intermediate 1

Intermediate 1 Step a

In a vial, 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (566 mg, 3.67mmol), methyl 2-bromobenzo[d]thiazole-6-carboxylate (500 mg, 1.837mmol), PdCl₂(dppf) DCM (75 mg, 0.092 mmol), and Na₂CO₃ (302 mg, 2.85mmol) were dissolved in dioxane (6.9 ml), and water (2.3 ml). Thereaction was heated to 100° C. for 3h. The reaction was cooled to roomtemperature and diluted with water. The aqueous layer was washed withEtOAc. The combined organic layer was dried over MgSO₄ and concentrated.The crude reaction mixture was purified by silica gel chromatographyeluting with 0-50% EtOAc/Hexanes to give the title compound (180 mg,0.821 mmol, 45%).

Intermediate 1 Steps b and c

In a vial, the compound from step a (120 mg, 0.547 mmol) was dissolvedin MeOH (5.47 ml). Pd/C (5.82 mg, 0.055 mmol) was added and the vial waspurged with H₂. Reaction allowed to stir at room temperature under H₂for 1 hr. Reaction purged with N₂ and reaction filtered through celite.Crude reaction mixture concentrated and purified by silica gelchromatography eluting with 0-50% EtOAc/Hexanes to give the titlecompound (80 mg, 0.362 mmol, 66%) The ester hydrolysis was carried outin a similar manner to Example 2 Step d. The title compound was isolatedby precipitation to afford the desired product (50 mg, 0.241 mmol, 67%).

Intermediate 2

In a vial, cyclopropyltrifluoro-14-borane, potassium salt (326 mg, 2.205mmol), methyl 2-bromobenzo[d]thiazole-6-carboxylate (200 mg, 0.735mmol), palladiumtetrakis (42.5 mg, 0.037 mmol) and potassium phosphate(468 mg, 2.205 mmol) were dissolved in Toluene (2.76 ml) and Water(0.919 ml). The reaction was heated to 100° C. and allowed to stirovernight. The reaction was cooled to room temperature and water wasadded. Aqueous layer washed with EtOAc. Combined organic layer driedover MgSO₄ and concentrated. Crude reaction mixture purified by silicagel chromatography eluting with 0-50% EtOAc/Hexanes to give the titlecompound (80 mg, 0.343 mmol, 47%).

The ester hydrolysis was carried out in a similar manner to Example 2Step d. The title compound was isolated by precipitation to afford thedesired product (50 mg, 0.228 mmol, 67%).

Intermediate 3

Intermediate 3 Step a

A solution methyl 2-amino-4-methoxybenzo[d]thiazole-6-carboxylate (2 g),CuBr₂ (3.7 g, 16.78 mmol) and t-BuNO₂ (1.7 g, 16.77 mmol) in CH₃CN wasstirred for 16 hours at room temperature under N₂ atmosphere. Theresulting solution was diluted with water, extracted with EtOAc and theorganic layer was dried, concentrated. The resulting solution waspurified by silica gel column chromatography (EtOAc in hexanes) toafford desired product (1.6 g, 63%) as orange solid. ESI-MS m/z: 301.90[M+H]⁺.

Intermediate 3 Step b

The title compound was synthesized in a similar manner to Intermediate 1using the compound from Intermediate 3 Step a. The ester hydrolysis wascarried out in a similar manner to Example 2 Step d. The title compoundwas isolated by precipitation to afford the desired product (30 mg,79%).

Intermediate 4

The title compound was synthesized in a similar manner to Intermediate 2using the compound from Intermediate 3 Step a. The ester hydrolysis wascarried out in a similar manner to Example 2 Step d. The title compoundwas isolated by precipitation to afford the desired product (57 mg,75%).

Intermediate 5

Intermediate 5 Step a

In a vial, methyl 6-amino-5-bromonicotinate (500 mg, 2.164 mmol) wasslurried in DCM (3.4 ml) and Pyridine (2.0 ml). The solution was cooledto 0° C. Cyclopropanecarbonyl chloride (590 μl, 6.49 mmol) was addeddropwise and the solution became homogenous. Reaction allowed to stirfor 2 hr. Reaction quenched upon addition of MeOH then concentrated.MeCN added and reaction evaporated to remove pyridine.

Dissolve in 1:1 THF/MeOH (5 mL) and cooled to 0° C. Added NaOMe in MeOH(620 μL, 2.7 mmol) slowly and allowed to stir 30 min. AcOH (250 uL)added which caused reaction to solidify. Solid broken up and wateradded. Solution allowed to stir a further 10 min before filtering andwashing solid with water to obtain title compound (546 mg, 1.825 mmol,84%).

Intermediate 5 Step b

In a vial, the compound from step a (400 mg, 1.337 mmol) was suspendedin THE (5.35 ml) and Lawesson's reagent (595 mg, 1.471 mmol) was added.The reaction was heated to 65° C. overnight. The reaction was allowed tocool to room temperature and stand before concentrating. Crude reactionmixture purified by silica gel chromatography eluting with 0-50%EtOAc/Hexanes to give the title compound (400 mg, 1.27 mmol, 95%).

Intermediate 5 steps c and d

In a vial, the compound from step b (434 mg, 1.377 mmol) was dissolvedin DMSO (3.44 ml). Sodium hydride (60.6 mg, 1.515 mmol) was added andthe reaction was allowed to stir 5 min before sealing vial and heatingto 70° C. for 5 h. Upon cooling, the reaction mixture was diluted withwater and extracted with EtOAc. The aqueous phase was washed with EtOAc.The combined organic phase was washed with brine, dried over MgSO₄,filtered, and concentrated. Crude reaction mixture purified by silicagel chromatography eluting with 0-50% EtOAc/Hexanes to give the titlecompound (130 mg, 0.555 mmol, 40%).

The ester hydrolysis was carried out in a similar manner to Example 2Step d. The title compound was isolated by precipitation to afford thedesired product (50 mg, 0.232 mmol, 77%).

Intermediate 6

Intermediate 6 Step a

A solution of 5-bromo-1H-indazole (500 mg, 2.54 mmol) in EtOAc (5 mL)were added 2,2-difluoro-2-(fluorosulfonyl)acetic acid (542 mg, 3.05mmol) and K₂CO₃ (701 mg, 5.08 mmol). The resulting mixture was stirredat rt for 2 hrs and then evaporated. The residue was purified byCombiflash eluting with 0-30% EtOAc/hexanes to obtain the desiredproduct (380 mg, 60.6%) as a white foam. ESI-MS m/z: 248.9 [M+H]⁺.

Intermediate 6 Steps b and c

To a solution of DMF/EtOH (6 mL, 2:1) were added compound from step a(240 mg, 0.866 mmol), Pd(OAc)₂ (23.34 mg, 0.104 mmol),1,3-Bis(diphenylphosphino)propane (86 mg, 0.208 mmol) and triethylamine(362 μl, 2.60 mmol). After degassed, CO balloon was subjected. Thereaction mixture was heated at 80° C. for 12 hrs, cooled down to rt anddiluted with EtOAc (150 mL). The solution was washed with brine, driedand purified by Combiflash eluting with 0-50% EtOAc/hexanes to obtainthe desired product (130 mg, 55.5%) was a white solid. ESI-MS m/z: 271.2[M+H]⁺.

To a solution of compound from step b (120 mg, 0.468 mmol) in THF/water(5:1, 6 mL) was added LiOH (112 mg, 4.68 mmol). The resulting mixturewas stirred at 45° C. for 20 hrs and then evaporated most of THF. Theremaining mixture was neutralized to pH ˜3 by adding aq. 1M HCl. Theprecipitated solid was filtered, washed with water, and dried to obtainthe desired product (105 mg, 93.0%) as a white solid. ESI-MS m/z: 243.21[M+H]⁺.

Intermediate 7 Steps a and b

To a solution of ethyl7-chloro-2-(difluoromethyl)-2H-indazole-5-carboxylate (200 mg, 0.728mmol) in 1,4-dioxane/water (2.1 mL, 20:1) were added cyclopropylboronicacid (125 mg, 1.456 mmol), XPhos Pd G2 (28.6 mg, 0.036 mmol) and K₃PO₄(773 mg, 3.64 mmol). After degassed, the mixture was heated at 80° C.for 2 hrs and the reaction was completed. The reaction mixture wasdiluted with EtOAc (100 mL), washed with brine, dried and purified bycombiflash eluting with 0-30% EtOAc/hexanes to obtain the desiredproduct (93 mg, 45.6%) as a brown solid. ESI-MS m/z: 281.10 [M+H]⁺.

To a solution of compound from step a (90 mg, 0.321 mmol) in THF/water(5:1, 6 mL) was added LiOH (77 mg, 3.21 mmol). The resulting mixture wasstirred at 45° C. for 20 hrs and then evaporated most of THF. Theremaining mixture was neutralized to pH ˜3 by adding aq. 1M HCl. Theprecipitated solid was filtered, washed with water, and dried to obtainthe desired product (60 mg, 74.1%) as a brown solid. ESI-MS m/z: 253.20[M+H]⁺.

Intermediate 8

Intermediate 8 Step a

To a solution of 5-bromo-3-methoxypyridin-2-amine (400 mg, 1.970 mmol)in EtOH (6 mL) was added 3-bromo-1,1,1-trifluoropropan-2-one (752 mg,3.94 mmol). The resulting mixture was heated at 80° C. overnight. Afterevaporated the solvent, the residue was purified by Combiflash elutingwith 0-40% EtOAc/hexanes to obtain the title compound (300 mg, 51.6%) asa colorless oil. ESI-MS m/z: 296.20 [M+H]⁺.

Intermediate 8 Steps b and c

The title compound was made according to Intermediate 6 step. The crudematerial was purified by Combiflash eluting with 0-50% EtOAc/hexanes toobtain the desired product (200 mg, 68.2%) was a white solid. ESI-MSm/z: 289.17 [M+H]⁺.

To a solution of compound from step b (200 mg, 0.694 mmol) in THF/water(5:1, 6 mL) was added LiOH (166 mg, 6.94 mmol). The resulting mixturewas stirred at rt for 4 hrs and then evaporated most of THF. Theremaining mixture was neutralized to pH ˜3 by adding aq. 1M HCl. Theprecipitated solid was filtered, washed with water, and dried to obtainthe desired product (120 mg, 66.5%) as a white solid. ESI-MS m/z: 261.20[M+H]⁺.

Intermediate 9

Intermediate 9 Step a

To a solution of 5-bromo-3-methoxypyridin-2-amine (2.0 g, 8.75 mmol) inEtOH (30 mL) was added 2-bromo-1-cyclopropylethan-1-one (1.99 g, 11.82mmol). The resulting mixture was heated at 80° C. for 3 days. Afterevaporated the solvent, the residue was purified by Combiflash elutingwith 0-100% EtOAc/hexanes and then 0-5% MeOH/DCM to obtain the titlecompound (1.88 g, 85%) as a brown foam. ESI-MS m/z: 268.20 [M+H]⁺.

Example 671 Steps b and c

To a solution of step a (500 mg, 1.872 mol) from step a in DCM (10 mL)was slowly added BBr₃ (3.74 mL, 3.74 mmol). The resulting mixture wasstirred at rt for 20 hrs and then quenched with water (2 mL). Afterdiluted with DCM (50 mL), the organic layer was separated, dried andevaporated. The residue was purified by Gilson prep-HPLC (20-90%,MeCN/Water, 25 min) to afford the desired product (40 mg, 8.44%) as abrown foam. ESI-MS m/z: 254.10 [M+H]⁺.

To a solution of step b (40 mg, 0.158 mmol) from step b in DMF (2 mL)were added K₂CO₃ (43.7 mg, 0.316 mmol) and 2,2-difluoroethyl4-methylbenzenesulfonate (187 mg, 0.79 mmol). The resulting mixture washeated at 60° C. in a sealed vessel for 20 hrs. After evaporated thesolvent, the residue was purified by Combiflash eluting with 0-30%EtOAc/hexanes to obtain the desired product (35 mg, 85%) as apale-yellow foam. ESI-MS m/z: 318.99 [M+H]⁺.

Intermediate 9 steps d and e

To a solution of DMF/EtOH (6 mL, 2:1) were added compound from step c(35 mg, 0.11 mmol), Pd(OAc)₂ (2.97 mg, 0.013 mmol),1,3-Bis(diphenylphosphino)propane (10.92 mg, 0.026 mmol) andtriethylamine (46 μl, 0.33 mmol). After degassed, CO balloon wassubjected. The reaction mixture was heated at 80° C. for 20 hrs, cooleddown to rt and diluted with EtOAc (50 mL). The solution was washed withbrine, dried and purified by Combiflash eluting with 0-50% EtOAc/hexanesto obtain the desired product (17 mg, 49.6%) was a white solid. ESI-MSm/z: 311.10 [M+H]⁺.

To a solution of compound from step d (17 mg, 0.055 mmol) in THF/water(5:1, 1 mL) was added 2 N NaOH (0.2 mL). The resulting mixture wasstirred at rt for 40 hrs and then evaporated most of THF. The remainingmixture was neutralized to pH ˜3 by adding aq. 1M HCl. After extractedthe mixture with 10% MeOH/DCM (50 mL×3), the combined organic layerswere combined, dried and evaporated to obtain the desired product (7 mg,45.3%) as a white solid. ESI-MS m/z: 283.07 [M+H]⁺.

Intermediate 10

Intermediate 10 Step a

To a solution of 2-amino-5-bromo-3-methoxybenzoic acid (4.00 g, 16.25mmol) in THE (60 mL) were added BH₃-THF (325 mL, 325.10 mmol) dropwiseunder ice/water bath and the reaction mixture was stirred at 50° C.overnight. The mixture was allowed to cool down to 0° C., quenched withMeOH and concentrated. The residue was diluted with aqueous Na₂CO₃ andextracted with EA. The combined organic layers were dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure to afford the crude product (4.4 g) as a yellow oil. ESI-MSm/z: 231.95 [M+H]⁺.

Intermediate 10 Steps b and c

A mixture of the compound from step a (4.40 g, 18.96 mmol) and MnO₂(8.24 g, 94.80 mmol) in DCM (50 mL) was stirred for overnight at roomtemperature. The resulting mixture was filtered, the filter cake waswashed with DCM. The filtrate was concentrated under reduced pressure.The residue was purified by silica gel column chromatography, elutedwith PE/EA (10:1) to afford the desired product (2.9 g, 66%) as a redsolid. ESI-MS m/z: 229.95 [M+H]⁺.

To a solution of the compound from step b (2.90 g, 12.60 mmol) in HCl(6M) (30 mL) at 0° C. was added NaNO₂ (1.09 g, 15.80 mmol) in water (5mL). After 30 min, the ice bath was removed and the reaction stirred atroom temperature for 30 min. The solid was removed by filtration, themother liquor was cooled to 0° C. and treated with NaN₃ (0.82 g, 12.60mmol) in water (5 mL). The ice bath was removed and stirring wascontinued for 30 min. The resulting solid (3.8 g) was collected byfiltration.

Intermediate 10 Step d

The compound from step c (3.80 g, 14.84 mmol) and aminocyclopropane(1.27 g, 22.26 mmol) in toluene (50 mL) were treated with molecularsieves. The reaction was stirred at room temperature for 2 hours, thenheated at reflux for overnight. After cooling to room temperature, themixture was filtered over celite and the solution was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with PE/EA (1:1) to afford the desired product(2.4 g, 60%) as a brown oil. ESI-MS m/z: 267.05 [M+H]⁺.

Intermediate 10 steps e and f

To a solution of the compound from step d (2.40 g, 8.98 mmol) in EtOH(10 mL) was added DMF (10 mL), TEA (2.5 mL), Pd(OAc)₂ (0.40 g, 1.78mmol), dppp (1.48 g, 3.59 mmol) in a pressure tank. The mixture waspressurized to 15 atm with carbon monoxide at 100° C. for overnight. Thereaction mixture was cooled to room temperature, concentrated undervacuum. The residue was purified by silica gel column chromatography,eluted with PE/EA (2:1) to afford the desired product (2.2 g, 94%) as ayellow oil. ESI-MS m/z: 261.10 [M+H]⁺.

Into a 100 mL round-bottom flask were added the compound from step e(2.10 g, 8.07 mmol) and MeOH (30 mL) at room temperature. A solution ofLiOH (1.93 g, 80.59 mmol) in H₂O (10 mL) was added and stirred for 2hours at room temperature. The resulting mixture was concentrated underreduced pressure. The mixture was acidified to pH 5 with HCl (2 M aq.).The product was collected by filtration and washed with water (1.28 g)ESI-MS m/z: 233.05 [M+H]⁺.

Intermediate 11 Step a, b, and c

A solution of methyl 1H-indazole-5-carboxylate (3 g, 17.03 mmol) andF-TEDA-BF₄ (12 g, 34.06 mmol) in DMF was stirred for 16 hours at 80° C.under nitrogen atmosphere. The resulting solution was purified byreverse phase C18 column chromatography (CH₃CN/H₂O) to afford desiredproduct (1.5 g, 45%) as a yellow solid. ESI-MS m/z: 194.95[M+H]⁺.

A solution of the compound from step a (1.5 g, 7.72 mmol), CH₃I (2.2 g,15.45 mmol) and Cs₂CO₃ (7.6 g, 23.17 mmol) in DMF was stirred for 4hours at room temperature. The resulting solution was diluted withwater, extracted with EA, the organic layer was dried, concentrated. Theresulting solution was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to afford desired product (600 mg, 37%) as ayellow solid. ESI-MS m/z: 209.00 [M+H]⁺.

A solution of the compound from step b (600 mg, 2.88 mmol), LiOH (690mg, 28.82 mmol) in THF/H₂O=10:1 (11 mL) was stirred for 16 hours at roomtemperature. The resulting mixture was concentrated under vacuum. Theresulting solution was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to afford desired product (211.9 mg, 38%) asa light-yellow solid. ESI-MS m/z: 195.05 [M+H]⁺.

Intermediate 12

Intermediate 12 Step a

To a mixture of 4-amino-3-fluoro-5-methylbenzonitrile (1.60 g, 10.65mmol) in HCl (8M) (15 mL) was added NaNO₂ (0.77 g, 11.19 mmol) in H₂O (2mL) dropwise at −5° C. The reaction was stirred for 30 min,2-methyl-2-propanethiol (0.96 g, 10.64 mmol) in EtOH (5 mL) was added at0° C. and stirred for 1 h. The reaction was quenched with Water/Ice,extracted with EA. The combined organic layers were concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with PE/EA (20:1) to afford the desired product(2.7 g, 99%) as an orange oil. ESI-MS m/z: 252.00 [M+H]⁺.

Intermediate 12 Step b

A mixture of the compound from step a (2.70 g, 10.74 mmol) and t-BuOK(9.64 g, 85.91 mmol) in DMSO (25 mL) was stirred for 30 min at roomtemperature. The resulting mixture was diluted with water, extractedwith EA. The combined organic layers were concentrated under reducedpressure. The residue was purified by reverse flash chromatography toafford the desired product (940 mg, 54%) as a yellow solid.

Intermediate 12 Steps c and d

A mixture of the compound from step b (900 mg, 5.58 mmol), iodoethane(1.05 g, 6.7 mmol) and K₂CO₃ (1.54 g, 11.17 mmol) in DMF (8 mL) wasstirred for overnight at room temperature. The resulting mixture waspoured into water, extracted with EA. The combined organic layers wereconcentrated under reduced pressure. The residue was purified by reverseflash chromatography to afford the desired product (510 mg, 48%) as anoff-white solid.

Into a 100 mL round-bottom flask were added the compound from step c(510 mg, 2.69 mmol), KOH (2.27 g, 40.46 mmol), EtOH (21 mL) and H₂O (7mL) at room temperature. The resulting mixture was stirred for overnightat 80° C. The resulting mixture was concentrated under reduced pressure.The residue was acidified to pH 5 with HCl (2 M aq.). The product wascollected by filtration and washed with water to afford the titlecompound (545 mg). ESI-MS m/z: 209.15 [M+H]⁺.

Intermediate 13 Steps a and b

A mixture of the compound from Intermediate 12 step b (900 mg, 5.58mmol), K₂CO₃ (1.54 g, 11.14 mmol) and iodoethane (1.05 g, 6.70 mmol,) inDMF (8 mL) was stirred for overnight at room temperature. The resultingmixture was poured into water, extracted with EA. The combined organiclayers were concentrated under reduced pressure. The residue waspurified by reverse flash chromatography to afford the desired product(280 mg, 26%) as an off-white solid.

Into a 100 mL round-bottom flask were added the compound from step a(280 mg, 1.48 mmol), KOH (1.25 g, 22.28 mmol), EtOH (21 mL) and H₂O (7mL) at room temperature. The resulting mixture was stirred for overnightat 80° C. The resulting mixture was concentrated under reduced pressure.The residue was acidified to pH 5 with HCl (2 M aq.). The product wascollected by filtration and washed with water to afford the titlecompound (263 mg) ESI-MS m/z: 209.15 [M+H]⁺.

Intermediate 14

Intermediate 14 Step a

Into a 250 mL 3-necked round-bottom flask were added4-bromo-2-chloro-6-methylpyridine (4.5 g, 22 mmol) and THE (100 mL). Theflask as cooled to −60° C. and LDA (4.43 mL, 33 mmol) was added dropwiseand stirred for 30 minutes. N-methoxy-N-methylcyclopropanecarboxamide(4.2 g, 33 mmol) was then added. The resulting mixture was stirred for1h at −60° C. under nitrogen atmosphere. The reaction was monitored byTLC. The reaction was quenched with sat. NH₄Cl (aq.) and the aqueouslayer was extracted with EtOAc. The resulting mixture was concentratedunder reduced pressure. The residue was purified by reverse flashchromatography with the following conditions: column, C18 silica gel;mobile phase, MeCN in water, 10% to 50% gradient in 25 min; detector, UV254 nm to afford the desired compound (2.8 g, 47%) as N off-white solid.ESI-MS m/z: 273.95 [M+H]⁺.

Intermediate 14 Step b

Into a 100 mL round-bottom flask were added the compound from step a(2.8 g, 10 mmol), NH₂OH HCl (3.5 g, 51 mmol), NaOH (2 g, 51 mmol) andMeOH (30 mL) at room temperature. The resulting mixture was stirred forovernight at 60° C. under nitrogen atmosphere. The reaction wasmonitored by LCMS. The aqueous layer was extracted with EtOAc. Theresulting mixture was concentrated under reduced pressure. The residuewas purified by silica gel column chromatography, eluted with PE/EtOAc(1:1) to afford the desired compound (2.2 g, 75%) as an off-white solid.ESI-MS m/z: 288.95 [M+H]⁺.

Intermediate 14 Steps c and d

Into a 100 mL round-bottom flask were added the compound from step b(2.2 g, 8 mmol), TFAA (1.8 g, 8 mmol), TEA (3.8 g, 38 mmol) and DME (15mL) at room temperature. The resulting mixture was stirred for 2h atroom temperature under nitrogen atmosphere. The reaction was monitoredby LCMS. Desired product could be detected by LCMS. The resultingmixture was concentrated under reduced pressure. The residue waspurified by reverse flash chromatography with the following conditions:column, C18 silica gel; mobile phase, MeCN in water, 10% to 50% gradientin 25 min; detector, UV 254 nm to afford the desired compound (1.8 g,87%) as off-white solid. ESI-MS m/z: 270.95 [M+H]⁺.

Into a 40 mL vial were added the compound from step c (1.8 g, 7 mmol),ferrous chloride (84 mg, 0.7 mmol) and DME (10 mL) at room temperature.The resulting mixture was stirred for 2h at 80° C. under nitrogenatmosphere. The reaction was monitored by TLC. The aqueous layer wasextracted with EtOAc. The resulting mixture was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with PE/EtOAc (1:1) to afford the desiredcompound (1.2 g, 67%) as an off-white solid. ESI-MS m/z: 270.95 [M+H]⁺.

Intermediate 14 Step e

Into a 40 mL vial were added the compound form step d (1.2 g, 4.4 mmol),MeONa (716 mg, 13 mmol) and THE (10 mL) at room temperature. Theresulting mixture was stirred for 2h at room temperature under nitrogenatmosphere. The reaction was monitored by LCMS. Desired product could bedetected by LCMS. The resulting mixture was concentrated under reducedpressure. The residue was purified by reverse flash chromatography withthe following conditions: column, C18 silica gel; mobile phase, MeCN inwater, 10% to 50% gradient in 25 min; detector, UV 254 nm to afford thedesired compound (800 mg, 68%) as off-white solid. ESI-MS m/z: 267.00[M+H]⁺.

Intermediate 14 Steps f and g

Into a 30 mL pressure tank reactor were added the compound form step e(400 mg, 1.5 mmol), Pd(AcO)₂ (101 mg, 0.45 mmol), DPPP (371 mg, 0.9mmol), EtOH (4 mL), Et₃N (1 mL) and DMF (4 mL) at room temperature. Theresulting mixture was stirred for overnight at 100° C. under COatmosphere at 10 atm. The reaction was monitored by TLC. The resultingmixture was filtered, the filter cake was washed with EtOH. The filtratewas concentrated under reduced pressure. The residue was purified byreverse flash chromatography with the following conditions: column, C18silica gel; mobile phase, MeCN in water, 10% to 70% gradient in 25 min;detector, UV 254 nm to afford the desired compound (360 mg, 92%) aswhite solid. ESI-MS m/z: 261.10 [M+H]⁺.

Into a 50 mL round-bottom flask were added the compound form step f (360mg, 1.4 mmol), LiOH (166 mg, 7 mmol), MeOH (5 mL) and H₂O (1 mL) at roomtemperature. The resulting mixture was stirred for 2h at roomtemperature under nitrogen atmosphere. The reaction was monitored byLCMS. The mixture was acidified to pH 6 with HCl (aq). The resultingmixture was concentrated under reduced pressure. The precipitated solidswere collected by filtration and washed with water (3×3 mL) to affordthe desired compound (280 mg, 87%) as off-white solid. ESI-MS m/z:233.00 [M+H]⁺.

Intermediate 15

Intermediate 15 Step a

A solution of 5-bromo-3-methoxypyridin-2-amine (5 g, 24.63 mmol) and2-bromo-1-cyclopropylethanone (8.03 g, 49.26 mmol) in EtOH (20 mL) wasstirred for overnight at 80° C. The residue was purified by reversephase flash to afford the desired product (4.4 g, 67%) as a yellowsolid. ESI-MS m/z: 267.00 [M+H]⁺.

Intermediate 15 steps b and c

A solution of the compound from step a (4.4 g, 16.47 mmol), Pd(OAc)₂(740 mg, 3.29 mmol) and dppp (2.7 g, 6.59 mmol) in DMF (12 mL), EtOH (12mL) and TEA (3 mL) was stirred overnight at 100° C. under CO atmosphere.The residue was purified by silica gel column chromatography, elutedwith PE/EA to afford the desired product (3 g, 70%) as a yellow solid.ESI-MS m/z: 261.00 [M+H]⁺.

A solution of the compound from step b (3 g, 11.53 mmol) and LiOH (2.76g, 115.25 mmol) in EtOH (10 mL) and H₂O (10 mL) was stirred for 2 hrs atroom temperature. The residue product was purified by reverse phaseflash to afford the desired product 1.3 g as a yellow solid. ESI-MS m/z:233.00 [M+H]⁺.

Intermediate 16

Intermediate 16 Step a

A solution of 5-bromo-3-methoxypyridin-2-amine (4 g, 19.7 mmol) andmethyl 3-bromo-2-oxopropanoate (7 g, 39.4 mmol) in EtOH (10 mL) wasstirred for 16 hrs at 80° C. The residue product was purified by reversephase flash to afford the desired product (4.6 g, 82%) as a yellowsolid. ESI-MS m/z: 285.00 [M+H]⁺.

Intermediate 16 steps b and c

A solution of the compound from step a (3 g, 10.5 mmol) and DIBAL-H (30mL) in THE (30 mL) was stirred for 1 hr at 0° C. The residue waspurified by silica gel column chromatography, eluted with DCM/MeOH(10/1) to afford the desired product (1.6 g, 59%) as a yellow solid.ESI-MS m/z: 257.00 [M+H]⁺.

A solution of the compound from step b (1 g, 3.9 mmol) and MnO₂ (3.4 g,38.9 mmol) in DCM (20 mL) was stirred overnight at room temperature. Theresulting mixture was filtered, and the filtrate was concentrated underreduced pressure. The crude 600 mg was used directly for next step.ESI-MS m/z: 255.00 [M+H]⁺.

Intermediate 16 Step d

A solution of the compound from step c (600 mg, 2.4 mmol) and DAST (1mL) in DCM (10 mL) was stirred for 2 hr at 0° C. The aqueous layer wasextracted with EA. The residue was purified by silica gel columnchromatography, eluted with PE/EA to afford the desired product (400 mg,61%) as a yellow solid. ESI-MS m/z: 277.00 [M+H]⁺.

Intermediate 16 Step e

A solution of the compound from step d (400 mg, 1.4 mmol) andPd(dppf)Cl₂ (211 mg, 0.29 mmol) in DMF (10 mL), H₂O (10 mL) and TEA (2mL) was stirred for 8 hrs at 100° C. under CO (15 atm). The residueproduct was purified by reverse phase flash to afford the desiredproduct (188 mg, 54%) as a yellow solid. ESI-MS m/z: 243.00 [M+H]⁺.

Intermediate 17

Intermediate 17 Step a

To a stirred solution of methyl 4-amino-3-methoxyphenyl carboxylate (500mg, 2.76 mmol) and 2-bromocyclopent-1-ene-1-carbaldehyde (1.45 g) intoluene were added BINAP (687 mg, 1.10 mmol) and Pd(OAc)₂ (124 mg, 0.55mmol). The reaction was heated to 90° C. under nitrogen atmosphere for 3h. The resulting mixture was extracted with EtOAc (3×0 mL). The combinedorganic layers were washed with EtOAc (3×10 mL), dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure. The residue was purified by silica gel column chromatography,eluted with PE/EtOAc (1:1) to afford the desired product (600 mg, 79%)as a yellow oil. ESI-MS m/z: 276.00 [M+H]⁺.

Intermediate 17 Steps b and c

A solution of the compound from step a (500 mg) and In(TfO)₂ (3 g) inxylene (5 mL) was stirred for 12 hr at 130° C. under N₂ atmosphere. Theresulting mixture was concentrated under reduced pressure for next stepdirectly. ESI-MS m/z: 258.00 [M+H]⁺.

A solution of the compound from step c (1 g, 3.89 mmol) and LiOH (93 mg,3.89 mmol) in H₂O (10 mL) and MeOH (10 mL) was stirred for 12 hr at roomtemperature. The crude product was purified by reverse phase flash toafford the desired product (184.7 mg) as a white solid. ESI-MS m/z:244.00 [M+H]⁺.

Intermediate 18

Intermediate 18 Steps a and b

A solution of (E)-2-bromocrotonaldehyde (10 g crude) and methyl4-amino-3-methoxybenzoate (34 g, 134.24 mmol) in AcOH:HCl (2:3, 50 mL)was stirred for 1 h at 100° C. under nitrogen atmosphere. The resultingsolution was concentrated to afford desired product (crude) as a yellowsolid. ESI-MS m/z: 295.90 [M+H]⁺.

A solution of the compound from step a, ethyl iodide (21 g, 135.08 mmol)and Cs₂CO₃ (66 g, 202.62 mmol) in DMF was stirred for overnight at roomtemperature. The resulting solution was diluted with water, extractedwith EA(×3), the organic layer was dried, concentrated. The resultingsolution was purified by silica gel column chromatography (PE:EA) toafford desired product (1 g) as a yellow solid. ESI-MS m/z: 323.90[M+H]⁺.

Intermediate 18 Steps c and d

A solution of the compound from step b (1 g, 3.08 mmol),tricyclohexylphosphine (0.9 g, 3.08 mmol), cyclopropylboronic acid (0.8g, 9.25 mmol), Pd(OAc)₂ (140 mg, 0.62 mmol) and K₃PO₄ (2 g, 9.25 mmol)in toluene and H₂O was stirred for overnight at 110° C. under nitrogenatmosphere. The resulting mixture was concentrated under vacuum. Theresulting solution was purified by silica gel column chromatography(PE:EA) to afford desired product as a yellow solid. ESI-MS m/z: 286.05[M+H]⁺.

A solution of the compound from step c and LiOH (441 mg, 18.43 mmol) inMeOH (10 mL) and H₂O (10 mL) was stirred for 1 h at room temperature.The resulting mixture was concentrated under vacuum. The resultingsolution was purified by reverse phase C18 column chromatography(CH3CN/H2O) to afford desired product (428 mg) as a yellow solid. ESI-MSm/z: 258.10 [M+H]⁺.

Intermediate 19 Steps a and b

A solution of Intermediate 18 Step b (200 mg, 0.62 mmol), L-proline (142mg, 1.23 mmol), chlorotrimethylaminyl (117 mg, 1.23 mmol) and Cu₂O (176mg, 1.23 mmol) in EtOH was stirred for 16 hours at 80° C. The resultingsolution was diluted with water, extracted with EA (×3), the organiclayer was dried, concentrated. The resulting solution was purified bysilica gel column chromatography (PE:EA) to afford desired product (70mg, 41%) as a yellow solid. ESI-MS m/z: 279.95 [M+H]⁺.

A solution of the compound from step a (70 mg, 0.25 mmol) and LiOH (60mg, 2.50 mmol) in THE (10 mL) and H₂O (1 mL) was stirred for 1 h at roomtemperature. The resulting mixture was concentrated under vacuum. Theresulting solution was purified by reverse phase C18 columnchromatography (MeCN/H₂O) to afford desired product (50 mg, 79%) as ayellow solid. ESI-MS m/z: 251.95 [M+H]⁺.

Intermediate 20 Steps a and b

A solution of Intermediate 18 Step b (200 mg, 0.62 mmol), methyl2,2-difluoro-2-sulfoacetate (237 mg, 1.23 mmol), KF (72 mg, 1.23 mmol)and CuI (235 mg, 1.23 mmol) in NMP was stirred for 4 hours at 120° C.The resulting solution was diluted with water, extracted with EA (×3),the organic layer was dried, concentrated. The resulting solution waspurified by silica gel column chromatography (PE:EA) to afford desiredproduct (140 mg, 72%) as a yellow solid. ESI-MS m/z: 313.95 [M+H]⁺.

A solution of the compound from step a (140 mg, 0.45 mmol) and LiOH (107mg, 4.47 mmol) in MeOH (10 mL) and H₂O (1 mL) was stirred for 1 h atroom temperature. The resulting mixture was concentrated under vacuum.The resulting solution was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to afford desired product (70 mg, 55%) as awhite solid. ESI-MS m/z: 285.90 [M+H]⁺.

Intermediate 21

Intermediate 21 Step a

To a stirred solution of methyl 4-amino-3-iodobenzoate (2.7 g, 10 mmol)in HCl (6 mL) were added NaNO₂ (0.7 g in 5 mL water) dropwise at 5° C.for 1 hr. To the above mixture was added piperidine (1 mL) dropwise at5° C. The resulting mixture was stirred for additional 1 hr at roomtemperature. The resulting mixture was extracted with EA and thecombined organic layers were washed with water, dried over anhydrousNa₂SO₄. The residue purified by silica gel column chromatography (EtOAcin hexanes) to afford the desired product (2.7 g) as a yellow solid.ESI-MS m/z: 374.00 [M+H]⁺.

Intermediate 21 Step b

To a dry and N₂-flushed 50 mL Schlenk tube, equipped with a magneticstirrer and a septum, was added bromo(prop-1-yn-1-yl)magnesium (4.3 g,29.94 mmol). The solution was cooled to −30° C. and ZnBr₂ (5.08 g, 22.56mmol) was added dropwise to the reaction mixture. The reaction mixturewas warmed to room temperature for 30 minutes. The compound from step a(2 g, 5.36 mmol) was added followed by (PPh₃)₄ (309 mg, 0.27 mmol). Thereaction mixture was stirred at room temperature for 2 hr and quenchedby saturated aqueous NH₄Cl. The aqueous was extracted with EtOAc, dried,and concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (EtOAc in hexanes) to afford thedesired product (2 g, 97%) as a yellow solid. ESI-MS m/z: 286.00 [M+H]⁺.

Intermediate 21 Step c

A solution of the compound from step b (1.5 g, 5.26 mmol,) and HBr inwater (850 mg, 10.51 mmol) in acetone (10 mL) was stirred for 2 hr atroom temperature. The resulting mixture was extracted with EtOAc and thecombined organic layers were washed with water and dried over anhydrousNa₂SO₄. The residue was purified by silica gel column chromatography(EtOAc in hexanes) to afford the desired product (900 mg, 61%) as ayellow solid. ESI-MS m/z: 281.00 [M+H]⁺.

Intermediate 21 Step d

A solution of the compound from step c (900 mg, 3.2 mmol) and Pd/C (681mg, 6.40 mmol) in MeOH (20 mL) was stirred for 2 hr at room temperatureunder H₂ atmosphere. The resulting mixture was filtered and the solutionwas concentrated to use directly for next step. ESI-MS m/z: 205.00[M+H]⁺.

Intermediate 21 Steps e and f

A solution of compound from step d and MnO₂ (1.5 g, 17.67 mmol) in THE(20 mL) was stirred for 2 hr at room temperature. The crude product waspurified by reverse phase flash to afford the desired product (253 mg,51%) as a yellow solid. ESI-MS m/z: 203.00 [M+H]⁺. In a vial, compoundfrom step e (100 mg, 0.495 mmol) and lithium hydroxide (118 mg, 4.95mmol) were dissolved in THF (2.2 ml), MeOH (2.2 ml), and Water (0.55ml). The reaction was allowed to stir at room temperature for 4 hours.Reaction diluted with water and the pH adjusted to 3-4 with 1M aq. HCl.Aqueous layer washed with DCM and 9:1 DCM/MeOH. Combined organic layerdried over MgSO₄ and concentrated under reduced pressure to furnish thetitle compound (45 mg, 48%). ESI-MS m/z: 188.68 [M+H]⁺.

Intermediate 21a

Intermediate 21a Step a

The following example was prepared in an analogous procedure toIntermediate 21 Step a with methyl 4-amino-3-iodo-5-methoxybenzoate (4g, 13.03) to afford the title compound. (3.5 g) ESI-MS m/z: 404.00[M+H]⁺.

Intermediate 21a Step b

The following example was prepared in an analogous procedure toIntermediate 21 Step b using cyclopropyl acetylene. The title compoundwas purified by silica gel chromatography to afford the title compoundas a brown oil (3.6 g). ESI-MS m/z: 342.00 [M+H]⁺.

Intermediate 21a Step c

The following example was prepared in an analogous procedure toIntermediate 21 Step c to afford the title compound (600 mg). ESI-MSm/z: 336.00 [M+H]⁺.

Intermediate 21a Step d

The following example was prepared in an analogous procedure toIntermediate 21 Step d to afford the title compound (500 mg). ESI-MSm/z: 261.00 [M+H]⁺.

Intermediate 21a Steps e and f

The following example was prepared in an analogous procedure toIntermediate 21 steps e and f to afford the title compound (200 mg,60%). ESI-MS m/z: 245.00 [M+H]⁺.

Intermediate 22

Intermediate 22 Step a

To a stirred solution of 2-amino-5-bromo-3-methoxybenzaldehyde (2.2 g,9.56 mmol), piperazine (2.5 g, 28.69 mmol) and methyl acetoacetate (2.2g, 19.13 mmol) in MeOH was stirred for 1 h at room temperature. Theresulting solution was diluted with water, extracted with EA(×3), theorganic layer was dried and concentrated. The resulting solution waspurified by silica gel column chromatography (PE:EA) to afford desiredproduct (2.5 g, 84%) as a yellow solid. ESI-MS m/z: 309.85 [M+H]⁺.

Intermediate 22 Steps b, c and d

A solution of the compound from step a (1 g, 28.69 mmol) in THE wastreated with DIBAL-H (20 mL) for 1 h at room temperature under nitrogenatmosphere. The resulting solution was diluted with water, extractedwith EA (×3), the organic layer was dried, concentrated. The resultingsolution was purified by silica gel column chromatography (PE:EA) toafford desired product (1 g crude) as a yellow solid. ESI-MS m/z: 281.90[M+H]⁺.

A solution of the compound from step b (1 g crude), Pd(OAc)₂ (159 mg,0.71 mmol), DPPP (585 mg, 1.42 mmol) and TEA (2.00 mL) in EtOH and DMFwas stirred for 16 hours at 100° C. under CO atmosphere (15 atm). Theresulting mixture was concentrated under vacuum. The resulting solutionwas purified by silica gel column chromatography (PE:EA) to afforddesired product (600 mg crude) as a yellow solid. ESI-MS m/z: 276.10[M+H]⁺.

A solution of the compound from step c (600 mg crude) and LiOH (522 mg,21.79 mmol) in MeOH/H₂O=10:1 (11 mL) was stirred for 1 hour at roomtemperature. The resulting mixture was concentrated under vacuum. Theresulting solution was purified by reverse phase C18 columnchromatography (MeCN/H₂O) to afford desired product (504.7 mg) as alight-yellow solid. ESI-MS m/z: 247.95 [M+H]⁺.

Intermediate 23

Intermediate 23 Step a

A solution of methyl 4-amino-3-hydroxybenzoate (5 g, 29.91 mmol) and NCS(4.8 g, 35.89 mmol) in CAN (20 mL) was stirred for 5 hr at 80° C. Theresidue was purified by silica gel column chromatography, eluted withPE/EA (10%) to afford the desired product (1.2 g) as a yellow solid.ESI-MS m/z: 202.00 [M+H]⁺.

Intermediate 23 Step b and c

A solution of the compound from step a (1.2 g, 5.95 mmol) and BrCN (3.2g, 29.74 mmol) in MeOH (42 mL) and H₂O (18 mL) was stirred for 48 hr at50° C. The resulting mixture was filtered, the filter cake was washedwith EA and water to get the desired product (700 mg, 52%). ESI-MS m/z:227.00 [M+H]⁺.

A solution of the compound from step b (500 mg, 2.21 mmol) and LiOH (264mg, 11.03 mmol) in THE (10 mL) and H₂O (10 mL) was stirred for 1 hr atroom temperature. The mixture was acidified to pH 7 with HCl (1M). Thecrude product was re-crystallized from the solution to afford thedesired product (231 mg, 49%) as a brown solid. ESI-MS m/z: 213.00[M+H]⁺.

Intermediate 24

Intermediate 24 Step a

A mixture of 5-bromo-3-chlorobenzene-1,2-diamine (2.5 g, 11.29 mmol),cyclopropanecarboxylic acid (1.26 g, 14.64 mmol), HATU (4.29 g, 11.29mmol), DIEA (2.9 g, 22.58 mmol) and DMF (50 mL) was stirred forovernight at room temperature. The reaction was monitored by LCMS. Theresulting mixture was poured into water, extracted with EtOAc. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure to afford the desired product (6.7 g, crude) as a brown oil.ESI-MS m/z: 289.00 [M+H]⁺.

Intermediate 24 Steps b and c

A solution of the compound from step a (6.7 g, 23.14 mmol) and aceticacid (60 mL) was stirred overnight at 100° C. The resulting mixture wasconcentrated under vacuum. The residue was purified by reverse flashchromatography with the following conditions: column, C18 silica gel;mobile phase, MeCN in water, 0% to 100% gradient in 30 min; detector, UV254 nm. to afford the desired product (2 g, 32%) as a yellow solid.ESI-MS m/z: 271.00 [M+H]⁺.

To a stirred solution of the compound from step b (2 g, 7.37 mmol) inDMF (20 mL) was added NaH (0.35 g, 14.73 mmol) in portions at 0° C.under nitrogen atmosphere. The resulting mixture was stirred for 30 minat the same temperature under nitrogen atmosphere. SEM-Cl (2.46 g, 14.76mmol) was added dropwise and stirred for 2 hr at room temperature. Thereaction was quenched with Water/Ice at room temperature. The resultingmixture was extracted with EtOAc. The combined organic layers werewashed with brine, dried over anhydrous Na₂SO₄. After filtration, thefiltrate was concentrated under reduced pressure to afford the desiredproduct (2.6 g, 88%) as a yellow oil. ESI-MS m/z: 401.00 [M+H]⁺.

Intermediate 24 Steps d and e

To a solution of the compound from step c (2.6 g, 6.47 mmol) in DMF (10mL), H₂O (10 mL) and TEA (2.5 mL) was added DPPP (1 g, 2.59 mmol) andPd(OAc)₂ (0.29 g, 1.29 mmol) in a pressure tank. The mixture waspressurized to 15 atm with carbon monoxide and stirred at 100° C. for 2days. The reaction mixture was cooled to room temperature and filteredto remove insoluble solids. The resulting mixture was concentrated underreduced pressure. The residue was purified by reverse flashchromatography with the following conditions: column, C18 silica gel;mobile phase, MeCN in water (0.05% FA), 10% to 80% gradient in 25 min;detector, UV 254 nm. to afford crude desired product (1.6 g, 67%) as ayellow solid. ESI-MS m/z: 367.10 [M+H]⁺.

A solution of the compound from step d (1.6 g, 4.4 mmol) and HCl (6M, 5mL) in EtOH (6 mL) and H₂O (6 mL) was stirred for 16 hr at 80° C. Thecrude product was purified by reverse phase flash to afford the desiredproduct (300 mg, 29%) as a white solid. ESI-MS m/z: 237.00 [M+H]⁺.

Intermediate 25 Steps a and b

A solution of lithium chloride (0.5 M solution in THF) (10.00 ml, 5.00mmol) and methyl-d3-magnesium iodide (1.0 M in diethyl ether) (5.00 ml,5.00 mmol) was treated with zinc chloride (1.9 M in 2-MeTHF) (1.316 ml,2.500 mmol) at room temperature. A solution of IPr PEPPSI (0.017 g,0.025 mmol) and methyl 7-bromo-1H-indazole-5-carboxylate (0.128 g, 0.500mmol) dissolved in 1.5 mL NMP was added dropwise and the resultingreaction mixture was stirred at room temperature overnight. Uponcompletion, the reaction was poured into 5% aq. citric acid andextracted with EtOAc. The organic extract was washed with brine, driedover anhydrous sodium sulfate, filtered, and concentrated. Purificationby flash column chromatography on silica gel afforded the title compound(84 mg, 0.435 mmol, 87% yield) as an off-white solid. ESI-MS m/z: 193.7[M+H]⁺.

A solution of step a (0.083 g, 0.430 mmol) in THF (5 ml) was treatedwith potassium trimethylsilanolate (0.276 g, 2.148 mmol) and stirred atroom temperature. Upon completion, the reaction was quenched with MeOHand silica gel, concentrated, and the resulting free-flowing admixturewas directly purified by flash column chromatography on silica gel toafford the title compound as a white solid (0.075 g, 97% yield). ESI-MSm/z: 179.6 [M+H]⁺.

Intermediate 26

Intermediate 26 Step a

Selenium dioxide (183 mg, 1.654 mmol) and2-methylbenzo[d]thiazole-6-carboxylic acid (213 mg, 1.102 mmol) weredissolved in 1,4-Dioxane (2.205 ml) at rt. The reaction mixture washeated at 90° C. for 16h. The mixture was diluted with ethyl acetate andpassed through a short silica gel plug. The plug was washed with ethylacetate and the resulting solution was concentrated and used as crude.ESI-MS m/z: 207.61 [M+H]⁺.

Intermediate 26 Step b

Step a (37 mg, 0.179 mmol) was dissolved in DCM (1 ml) and DAST (83 μl,0.625 mmol) was added at 0° C. The reaction mixture was allowed to stirat 0° C. for 2 h. The reaction mixture was diluted with ethyl acetateand poured into 1N HCl solution. The aqueous layer was extracted withethyl acetate for 3 times. The combined organic layers were dried andpurified by flash chromatography to give the title compound. (15 mg,36.6% yield). ESI-MS m/z: 230.00 [M+H]⁺.

Intermediate 27 Steps a and b

A solution of lithium chloride (0.5 M solution in THF) (6.17 ml, 3.08mmol) and methyl-d3-magnesium iodide (1.0 M in diethyl ether) (3.08 ml,3.08 mmol) was treated with zinc chloride (1.9 M in 2-MeTHF) (0.812 ml,1.542 mmol) at room temperature. A solution of IPr PEPPSI (10.51 mg,0.015 mmol) and ethyl 3-bromo-8-methoxy-2-methylquinoline-6-carboxylate(0.1 g, 0.308 mmol) dissolved in 1.5 mL NMP was added dropwise and theresulting reaction mixture was stirred at room temperature overnight.Upon completion, the reaction was poured into 5% aq. citric acid andextracted with EtOAc. The organic extract was washed with brine, driedover anhydrous sodium sulfate, filtered, and concentrated. Purificationby flash chromatography afforded the desired product (50 mg, 61.8%yield) as a yellow solid. ESI-MS m/z: 263.02 [M+H]⁺.

The ester hydrolysis was carried out according to Example 2 Step d toafford the desired compound.

Example 3

Example 3 Step a

To a solution of (R)-3-cyclopropoxy-4-(2-hydroxypropoxy)benzoic acid(100 mg, 0.396 mmol) in THE (1.982 ml) was added 4-methylmorpholine(47.9 μl, 0.436 mmol) and tert-butylchlorodimethylsilane (65.7 mg, 0.436mmol). After stirring the reaction mixture for 30 min, 1H-tetrazole(4405 μl, 1.982 mmol) and dibenzyl diisopropylphosphoramidite (400 μl,1.189 mmol) were added. The reaction mixture was allowed to stir at roomtemperature for 5 h. The reaction mixture was cooled to 0° C., and thenhydrogen peroxide (121 μl, 1.189 mmol) was added. The reaction mixturewas slowly warmed to room temperature over 30 min. Saturated sodiumsulfite was then added. After stirring vigorously for 30 min, themixture was partitioned between ethyl acetate and saturated sodiumsulfite. The pH value of the combined aqueous layers was adjusted to 3with 1N HCl, and the resulting suspension was extracted with ethylacetate. The combined organic layers were dried over MgSO₄. The solventwas removed, and the residue was purified by flash chromatography toyield the title compound (183 mg, 90% yield).

Example 3 Steps b and c

The amide coupling was carried out according to Example 1 Step c toafford the desired compound.

Step b (193 mg, 0.216 mmol) was dissolved in Ethyl acetate (2.159 ml) ina 40 mL vial. The vial was flushed with nitrogen gas three times.palladium on carbon (22.98 mg, 0.022 mmol) was added in one portion. Thevial was flushed with hydrogen balloon and the reaction mixture wasallowed to stir at 25° C. for 2h. The hydrogen gas was removed andflushed with nitrogen. The solution was filtered through celite and theresidue was concentrated and purified by HPLC. ESI-MS m/z: 714.13[M+H]⁺.

Example 4

Example 4 Step a

The following example was prepared according to Example 1 Step c withacid coupling partner 2-formyl-8-methoxy-3-methylquinoline-6-carboxylicacid (55 mg).

Example 4 Step b

The product from Step a (36 mg, 0.057 mmol) was dissolved in EtOH andsodium borohydride (4.35 mg, 0.115 mmol) was added at 0° C. The reactionmixture was allowed to stir at rt for 30 min. The reaction mixture wasquenched by adding saturated NH₄Cl carefully. The aqueous layer wasextracted with ethyl acetate and the combined organic layers were driedand purified by HPLC to afford the title compound (14.5 mg, 40.1%yield). ESI-MS m/z: 629.27 [M+H]⁺.

Example 5

Then Example 4 Step a (38 mg, 0.061 mmol) was dissolved in tBuOH andwater. 2-methyl-2-butene (129 μl, 1.213 mmol) and sodium dihydrogenphosphate (72.8 mg, 0.606 mmol) were added at rt. Then sodium chlorite(68.6 mg, 0.606 mmol) was added. The reaction mixture was allowed tostir at rt for 30 min. The reaction was quenched by adding 15% Na₂S203solution and the aqueous layer was extracted with ethyl acetate. Thecombined organic layers were dried and purified by HPLC to give thetitle compound (14 mg, 0.022 mmol, 35.9% yield). ESI-MS m/z: 643.15[M+H]⁺.

Intermediate 28 Steps a and b

A 100 mL rbf with stirbar was charged with methyl4-amino-3-methoxybenzoate (2.86 g, 15.79 mmol) and (E)-2-ethylbut-2-enal(5.468 g, 18.94 mmol) then AcOH (23.68 ml) and HCl (15.79 ml). Themixture was then heated to 100° C. for 2 h. The mixture was then cooledto room temperature and partially concentrated at 60° C. under vacuum toafford a thick black oil. This oil was treated with 40 mL of EtOH and250 μL of sulfuric acid then heated to 80° C. for 2 h. The mixture wascooled to room temperature then neutralized by the addition of 2 mLNEt₃, and concentrated. The resulting residue was purified by purifiedby automated silica gel column chromatography (0 to 45% ethyl acetate inhexanes) and dried under high vacuum to give the title compound as acolorless oil (49.8 mg, 1%). ESI-MS m/z: 274.2 [M+H]⁺.

The ester hydrolysis was carried out in a similar manner to Example 2Step d. The crude product was carried forward without furtherpurification as a white solid. ESI-MS m/z: 246.0 [M+H]⁺.

Intermediate 29

A 4 mL vial was charged with 2-methyl-1H-indole-5-carboxylic acid (125mg, 0.714 mmol) then DMF (1.427 ml), the mixture was cooled to 0° C.then NaH (86 mg, 2.141 mmol) was added slowly. The mixture was stirredfor 15 min then methyiodide (134 μL, 2.141 mmol) was added. After 2 h,the mixture was neutralized by addition of 1 M HCl (aq) then dilutedwith 2 mL dichloromethane. The layers were separated and the aqueouslayer was washed with dichloromethane 5×1 mL. The combined organics werewashed with water, then dried over Na₂SO₄ filtered and concentrated thenpurified by automated silica gel column chromatography and dried underhigh vacuum to give the title compound as a white solid (65 mg, 38%).ESI-MS m/z: 189.7 [M+H]⁺.

Intermediate 30

Intermediate 30 Step a

A 20 mL vial was charged with 3-methyl-1H-pyrazol-5-amine (1.00 g, 10.3mmol) AcOH (20.6 ml) and diethyl but-2-ynedioate (1.65 ml, 10.3 mmol).The mixture was stirred at 23° C. for 48 h, then poured intohexanes:EtOAc (1:1, 100 mL) and stirred for 1 h. The solids werecollected by filtration to afford the title compound as a yellow solid(1.78 g, 78%) ESI-MS m/z: 221.9 [M+H]⁺.

Intermediate 30 Steps b and c

A 20 mL vial was charged with a 20 mL vial was charged with step a (250mg, 1.13 mmol) cesium carbonate (736 mg, 2.26 mmol) DMF (2.3 ml) thenMel (212 μl, 3.39 mmol) was added. The mixture was stirred for 16 h. Themixture was then diluted with 5 mL water, and 5 mL dichloromethane. Thelayers were separated, and the aqueous layer was washed 5×5 mL withdichloromethane. The combined organics were washed with water 2×1 mL,then brine 1 mL, dried over Na₂SO₄, concentrated and purified byautomated silica gel chromatography to afford the title compound as ayellow oil (205 mg, 77%) ESI-MS m/z: 235.9 [M+H]⁺. The ester hydrolysiswas carried out in a similar manner to Example 2 Step d. Afteracidification the product was collected by filtration and carriedforward without further purification as a yellow solid. ESI-MS m/z:207.7 [M+H]⁺.

Intermediate 31

A 20 mL vial was charged with 6-methylnicotinic acid (333 mg, 2.43 mmol)and 2-bromo-1-cyclopropylethan-1-one (396 mg, 2.43 mmol) thenacetonitrile (12.0 ml) and heated to 70° C. for 16 h, then triethylamine(1.35 mL, 9.71 mmol) was added and the mixture stirred an additional 3h. The mixture was then diluted with 5 mL water and 5 mL EtOAc. Thelayers were separated and the aqueous layer was washed with EtOAc 4×5mL. The combined organics were washed with 2 mL brine, dried overNa₂SO₄, filtered, concentrated and purifed by automated silica gelchromatography to afford the title compound (78 mg, 16%) ESI-MS m/z:201.8 [M+H]⁺.

Intermediate 32 steps a and b

An 8 mL vial was charged with6-bromo-2-cyclopropylimidazo[1,2-b]pyridazine (128 mg, 0.538 mmol)palladium(II) acetate (12 mg, 0.054 mmol)1,3-bis(diphenylphosphino)propane (44.3 mg, 0.108 mmol), and theheadspace of the vial was purged with N₂ for 30 min. A mixture of DMF(1.434 ml) ethanol (0.717 ml) and triethylamine (225 μl, 1.613 mmol) wasdegassed by sparging with N₂ for 30 min, then added to the mixture ofsolids. The vial was placed under a balloon of CO, and the mixture washeated to 80° C. for 16 h. Additional palladium(II) acetate (12 mg,0.054 mmol) was added and the mixture heated to 100° C. for 4 h. Thereaction vessel was degassed by sparging with N₂, then the mixture wasconcentrated and purified by automated silica gel chromatography toafford the title compound as a yellow oil (35 mg, 28%). ESI-MS m/z:232.0 [M+H]⁺.

The ester hydrolysis was carried out in a similar manner to Example 2Step d. The product could not be extracted from the aqueous layer. Theaqueous layer was concentrated to afford the title compound as a mixturewith lithium chloride. ESI-MS m/z: 204.1 [M+H]⁺.

Intermediate 33

Intermediate 33 Step a

A 20 mL vial was charged with 1H-pyrrol-1-amine (1 ml, 13.15 mmol) anddiethyl 2-(ethoxymethylene)malonate (3.11 ml, 15.39 mmol) they were,heated to 125° C. for 1 h. The mixture was diluted with a mixture ofbiphenyl (1 g) and diphenylether (3 ml) then heated to 200° C. for 2 h.The mixture was cooled to room temperature then purified by automatedsilica gel chromatography (0 to 10% ethyl acetate in cyclohexane) toafford the title compound as a yellow solid (1.43 g, 53%) ESI-MS m/z:207.1 [M+H]⁺.

Intermediate 33 Step b

An 8 mL vial was charged with the compound from step a (300 mg, 1.46mmol) CCl₄ (1.5 ml) celite (150 mg) and triphenylphosphine (1.13 g, 4.36mmol). The mixture was heated to 75° C. for 16 h. The reaction wasdiluted with EtOAc and filtered through celite. The filtrate wasconcentrated and purified by automated silica gel chromatography toafford the expected ester (243 mg, 74%) as well as the hydrolysisproduct as a fluorescent yellow solid (19 mg, 7%) which was carriedforward directly. ESI-MS m/z: 197.0/199.0 [M+H]⁺.

Intermediate 34

An 8 mL vial was charged with the ester from Intermediate 33 step b (131mg, 0.583 mmol) and ammonium formate (184 mg, 2.92 mmol). A mixture ofdioxane (2.92 ml) and ethanol (2.92 ml) was degassed by sparging with N2for 30 min, then added to the mixture. Palladium on carbon (62.1 mg,0.583 mmol, 10 wt %) was added and the mixture allowed to stir for 1 hthen filtered through celite. The filtrate was concentrated and purifiedby automated silica gel chromatography to afford the product as afluorescent yellow solid (77 mg, 70%) ESI-MS m/z: 191.1 [M+H]⁺.

The ester hydrolysis was carried out in a similar manner to Example 2Step d. After acidification the title compound was collected byfiltration as a fluorescent yellow solid (19 mg, 32%) ESI-MS m/z: 163.0[M+H]⁺.

Intermediate 35 Steps a and b

An 8 mL vial was charged with methyl 4-amino-3-bromobenzoate (345 mg,1.500 mmol) Pd(PPh₃)₂Cl₂ (211 mg, 0.300 mmol), copper(I) iodide (28.6mg, 0.150 mmol), the reaction vessel was evacuated and backfilled withN₂ three times. Toluene (2 mL) and water (1 mL) were degassed bysparging with N₂ for 15 min, then added to the vial with the solids.Ethynylcyclopropane (508 μL, 6.00 mmol) and triethylamine (627 μL, 4.50mmol) were added and the mixture stirred for 16 h at 70° C. The mixturewas then diluted with water and EtOAc, the phases were separated, theorganic layer was concentrated and purified by automated silica gelchromatography (0 to 10% ethyl acetate in cyclohexane). To afford thetitle compound as a colorless oil (298 mg, 92%) ESI-MS m/z: 216.1[M+H]⁺.

An 8 mL vial containing the compound from step a (99 mg, 0.46 mmol) wascharged with dimethylsulfoxide (1 ml), and KOtBu (155 mg, 1.38 mmol)then heated to 100° C. for 30 min. The mixture was cooled to roomtemperature and diluted with water and EtOAc, then acidified to pH 3with 1 M (aq) HCl. The layers were separated and the aqueous layerwashed 4×3 mL with EtOAc. The combined organic layers were then washedtwice with water, then with brine, dried over Na₂SO₄ and purified byautomated silica gel chromatography (0 to 50% (1% AcOH in EtOAc) incyclohexane) to afford the title compound as a white solid (45 mg, 49%).ESI-MS m/z: 202.1 [M+H]⁺.

Intermediate 36

Intermediate 36 Step a

Into a 100 mL round-bottom flask were added Example 1, step b (3.80 g,10.27 mmol), acetone (100 mL), the solution was cooled to 0° C., andthen Jones reagent (1.9-2.2 M, 10 mL) was added dropwise (with internaltemperature monitoring). The reaction was warmed to room temperature andmonitored by LCMS (3 hr). The reaction was cooled to 0° C., quenchedwith ^(i)PrOH and stirred for 15 minutes. The reaction was diluted withEtOAc and water. The aqueous was extracted, the combined organics weredried and concentrated under reduced pressure to get the crude productas a yellow solid (3.95 g, 99%). ESI-MS m/z: 383.80 [M+H]⁺.

Intermediate 36 Step b

Into a 100 mL round-bottom flask were added the compound from step a(3.95 g, 10.28 mmol), NH₄C₁ (1.10 g, 20.57 mmol) and the solidsdissolved in DMF (20 mL). Hunig's base (5.27 mL, 30.84 mmol) was added,the reaction was cooled to 0° C., and HATU (7.82 g, 20.56 mmol) wasadded. The reaction was warmed to room temperature and monitored by LCMS(1 hr). The reaction was diluted with EtOAc and water. The aqueous wasextracted, the combined organics were dried and concentrated. Thematerial was purified by automated column chromatography (silica gel,0-100% EtOAc in hexanes) to afford the title compound (3 g, 76%). ESI-MSm/z: 382.95 [M+H]⁺.

Intermediate 36 Step c

Into a 100 mL round-bottom flask were added the compound from step b(3.00 g, 7.83 mmol), 3,3,3-trifluoroprop-1-en-2-ylboronic acid (2.19 g,15.65 mmol), Pd(dppf)Cl₂ (1.15 g, 1.56 mmol), and the material dissolvedin dioxane (40 mL) and H₂O (5 mL). K₂CO₃ (3.25 g, 23.50 mmol) was thenadded and the resulting mixture was stirred for 1 h at 90° C. undernitrogen atmosphere. The mixture was cooled to room temperature, pouredinto water, extracted with EtOAc and the combined organics wereconcentrated under reduced pressure. The residue was purified by columnchromatography (silica gel, 0-100% EtOAc in hexanes) to afford desiredproduct as a brown oil (2.3 g, 83%). ESI-MS m/z: 350.90 [M+H]⁺.

Intermediate 36 Step d

To a stirred solution of step c (5.00 g, 14.24 mmol) and3-chloro-4-fluorophenylboronic acid (3.72 g, 21.33 mmol) in THE (80 mL)were added Na₂CO₃ (3.32 g, 31.33 mmol), H₂O (20 mL) and Pd(PPh₃)₂C₁₂(1.00 g, 1.42 mmol). The resulting mixture was stirred for 1 h at 70° C.under nitrogen atmosphere. The reaction was monitored by TLC and LCMS.The resulting mixture was extracted with EtOAc, and the combined organiclayers were washed with brine, dried, and concentrated under reducedpressure. The residue was purified by automated column chromatography(silica gel, 0-75% EtOAc in hexanes) to afford the title compound as ayellow solid (5.8 g, 99%). ESI-MS m/z: 401.05 [M+H]⁺.

Intermediate 36 Step e

To a 500 mL round-bottom flask equipped with a stir bar was addedAD-mix-β (33.82 g, 43.41 mmol) and methanesulfonamide (1.38 g, 14.47mmol). The solids were dissolved in tBuOH (60 mL) and H₂O (100 mL), andthe flask cooled to 0° C. and the compound from step d (5.80 g, 14.47mmol) was added slowly as a solution of tBuOH (40 mL). The reaction wasallowed to warm to room temperature naturally and stirred for 16 hrs.The reaction was quenched with the addition of sodium sulfite (0.25 gper g AD-mix), diluted with water and EtOAc. The layers were separated,and the aqueous layer was extracted with EtOAc. The combined organicswere washed with brine, dried over Na₂SO₄, filtered concentrated andpurified by automated column chromatography (silica gel, 0-100% EtOAc inhexanes) to afford the title compound as a white solid (5.48 g, 87%).ESI-MS m/z: 435. [M+H]⁺.

Intermediate 36 Step f

Into a 250 mL round-bottom flask were added the compound from step e(4.70 g, 10.81 mmol) and DCM (80 mL) at room temperature. The solutionwas cooled to 0° C., and DMAP (264 mg, 2.16 mmol), TEA (3.28 g, 32.43mmol and TsCl (2.47 g, 12.97 mmol) were then sequentially added. Theresulting mixture was stirred for 1 h at 0° C. The mixture was acidifiedto pH 4 with 2 M HCl, and the aqueous extracted with DCM. The combinedorganic layers were dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to afford the crude product as a light-yellow solid(6.2 g, 97%). ESI-MS m/z: 589.15 [M+H]⁺.

Intermediate 36 Step g

Into a 100 mL round-bottom flask was added NH₃ in MeOH (35 mL) at roomtemperature, and the compound from step g (6.20 g, 10.52 mmol) wasslowly added. The resulting mixture was stirred at room temperature andmonitored by LCMS (5 hr). The mixture was dissolved in EtOAc, washedwith sat. sodium bicarbonate 3×, brine, dried, and concentrated toafford the title compound (2.93 g, 64%). ESI-MS m/z: 434.05 [M+H]⁺.

Intermediate 37

Intermediate 37 Step a

To a 20 mL vial equipped with a stir bar was addedN-fluoro-N-(phenylsulfonyl)benzenesulfonamide (963 mg, 3.05 mmol) andmethyl 7-bromo-5-iodo-2,3-dihydrofuro[2,3-c]pyridine-3-carboxylate (586mg, 1.526 mmol), and the solids were dissolved in THE (0.25M). The vialwas cooled to −78° C., and LDA (916 μl, 1.831 mmol) was slowly added.The reaction was stirred at −78 C for 1 hr. After 1 hr at −78° C.,reaction gelled up. The reaction was warmed to rt and stirred andmonitored by LCMS. The reaction was diluted with EtOAc and quenched withwater and sat. ammonium chloride. EtOAc and DCM/MeOH 1× each extractionwith a phase separator cartridge. Concentrated. The material waspurified by automated column chromatography (0-100% EtOAc/cHex) toafford the title compound (483 mg, 71%). ESI-MS m/z: 401.86/403.86[M+H]⁺.

Intermediate 37 Step b

To a 20 mL vial containing step a (483 mg, 1.081 mmol) was added andstir bar, and the material was dissolved in THF/MeOH, and Water (1:1:1,0.25 M). Lithium hydroxide hydrate (113 mg, 2.70 mmol) was added, thereaction stirred at room temperature and monitored by LCMS. The reactionwas diluted with EtOAc and acidified to pH=2 with 2M HCl. EtOAc andDCM/MeOH extraction with a phase separator cartridge. The material wastritterated with DCM/hexanes to afford an orange solid. (455 mg, 92%)ESI-MS m/z: 387.82/387.82 [M+H]⁺.

Intermediate 37 Step c

The above compound was prepared in an analogous fashion to Intermediate36 Step b. The crude material was purified by automated columnchromatography (silica gel, 0-100% EtOAc/cHex) to afford the titlecompound as an orange oil (243 mg, 43%) ESI-MS m/z: 386.33 [M+H]⁺.

Intermediate 38

The above example was prepared in an analogous sequence to Intermediate36 as a mixture of diastereomers to afford the desired amino alcohol (42mg). ESI-MS m/z: 404.09 [M+H]⁺.

The compounds in Table 1 were prepared by a method similar to that ofExample 1 step c (PyBOP or HATU). The majority of compounds werepurified by Gilson prep-HPLC, and some were purified by automated columnchromatography (silica gel). Many of the aryl acid coupling partnerswere prepared according to Intermediates 1-38, or analogous procedures.In certain cases the aryl acid coupling partners were commerciallyavailable or prepared by previously described methods.

TABLE 1 MS⁺ Example Structure m/z  6

586.21  7

596.19  8

603.96  9

600.21  10

634.19  11

589.16  12

628.01  13

645.82  14

601.06  15

593.05  16

619.18  17

631.12  18

634.99  19

665.05  20

652.99  21

601.94  22

576.17  23

556.40  24

614.19  25

602.17  26

640.20  27

614.18  28

648.14  29

636.13  30

674.16  31

576.14  32

648.14  33

584.17  34

599.19  35

671.25  36

587.19  37

659.25  38

652.22  39

686.18  40

624.10  41

594.16  42

628.09  43

612.13  44

662.06  45

646.09  46

658.11  47

634.16  48

668.12  49

642.14  50

558.18  51

584.19  52

530.22  53

556.24  54

612.30  55

592.12  56

559.17  57

602.17  58

636.13  59

648.14  60

618.14  61

617.15  62

606.16  63

594.13  64

624.14  65

658.10  66

620.17  67

618.15  68

664.17  69

623.05  70

603.05  71

625.00  72

629.15  73

657.15  74

599.40  75

629.15  76

643.05  77

609.16  78

639.10  79

605.15  80

631.05  81

626.00  82

633.00  83

667.07  84

618.20  85

652.15  86

660.20  87

559.20  88

584.20  89

590.15  90

578.15  91

594.10  92

614.15  93

602.15  94

618.15  95

614.15  96

657.20  97

659.05  98

658.10  99

656.15 100

558.2 101

558.1 102

572.2 103

578.0 104

562.1 105

612.1 106

578.0 107

545.1 108

574.0 109

545.1 110

546.1 111

560.00 112

628.30 113

676.20 114

702.50 115

704.50 116

730.30 117

690.50 118

718.50 119

680.50 120

678.10 121

692.20 122

558.00 123

586.20 124

548.20 125

586.20 126

584.10 127

558.20 128

561.10 129

576.50 130

604.12 131

661.50 132

661.50 133

607.60 134

619.6 135

687.60 136

643.50 137

625.5 138

606.60 139

603.60 140

653.60 141

610.50 142

592.50 143

615.60 144

613.60 145

618.00 146

601.50 147

630.60 148

648.60 149

687.50 150

606.60 151

620.60 152

632.60 153

646.60 154

646.60 155

674.70 156

674.70 157

647.60 158

647.60 159

617.60 160

672.70 161

590.60 162

619.60 163

633.60 164

633.60 165

618.60 166

654.60 167

628.60 168

572.20 169

586.20 170

582.30 171

557.10 172

606.10 173

606.10 174

627.50 175

571.20 176

557.10 177

617.20 178

639.30 179

667.30 180

589.10 181

573.10 182

651.10 183

673.30 184

701.20 185

583.10 186

585.20 187

663.20 188

663.10 189

557.20 190

576.10 191

560.20 192

615.10 193

584.20 194

557.20 195

621.10 196

542.30 197

561.10 198

578.10 199

577.10 200

572.30 201

544.10 202

638.00 203

638.00 204

662.20 205

583.20 206

599.20 207

600.20 208

600.20 209

600.20 210

642.23 211

609.17 212

570.19 213

600.22 214

600.22 215

725.31 216

668.25 217

614.24 218

687.14 219

701.17 220

721.11 221

667.21 222

687.15 223

627.19 224

647.12 225

589.15 226

681.22 227

709.21 228

667.10 229

585.16 232

628.15

Intermediate 39

Intermediate 39 Step a

Into a 250 mL 3-necked round-bottom flask was added4-bromo-2-chloro-6-methylpyridine (3 g, 15 mmol), cyclopropanecarboxylicacid, methyl ester (22 mL, 22 mmol) and THF (50 mL) at 0° C. To theabove mixture was added LiHMDS (3.7 g, 22 mmol) dropwise. The resultingmixture was stirred for additional 2 hr at 0° C. The reaction wasmonitored by TLC. The reaction was quenched by the addition of sat.NH₄Cl (aq.). The aqueous layer was extracted with EtOAc. The resultingmixture was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography, eluted with PE/EA (7:1) toafford the desired compound (3.3 g, 83%) as a light yellow solid. ESI-MSm/z: 273.95 [M+H]⁺.

Intermediate 39 Step b

Into a 40 mL vial was added the compound from step a (3.3 g, 12 mmol),NaOAc (5 g, 60 mmol), hydroxylamine hydrochloride (4.2 g, 60 mmol) andMeOH (20 mL) at room temperature. The resulting mixture was stirred forovernight at 60° C. under nitrogen atmosphere. The reaction wasmonitored by LCMS. The aqueous layer was extracted with EtOAc. Theresulting mixture was concentrated under reduced pressure. The residuewas purified by silica gel column chromatography, eluted with PE/EA(5:1) to afford the desired compound (2.3 g, 66%) as a white solid.ESI-MS m/z: 288.95 [M+H]⁺.

Intermediate 39 Steps c and d

Into a 20 mL vial was added the compound from step b (1.1 g, 3.8 mmol)and DME (10 mL) at 0° C. To the above mixture was added TFAA (0.9 g, 4mmol) dropwise. The resulting mixture was stirred for additional 3 hr atroom temperature. The reaction was monitored by TLC. The residue waspurified by reverse flash chromatography with MeCN in water to give thedesired compound (600 mg, 58%) as white solid. ESI-MS m/z: 270.95[M+H]⁺.

Into a 20 mL vial was added the compound from step c (600 mg, 2.2 mmol),ferrous chloride (56 mg, 0.44 mmol) and DME (10 mL) at room temperature.The resulting mixture was stirred for 2h at 80° C. under nitrogenatmosphere. The reaction was monitored by TLC. The aqueous layer wasextracted with EtOAc. The resulting mixture was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with PE/EA (5:1) to afford the desired compound(400 mg, 67%) as a white solid. ESI-MS m/z: 270.95 [M+H]⁺.

Intermediate 39 Step e

Into a 50 mL round-bottom flask was added the compound from step d (400mg, 1.5 mmol), cyclopropanol (257 mg, 4.4 mmol), NaH (71 mg, 3 mmol) andDMF (10 mL) at 0° C. The resulting mixture was stirred for 2 hr at roomtemperature under nitrogen atmosphere. The reaction was quenched by theaddition of sat. NH₄Cl (aq.). The aqueous layer was extracted with EtOAcand concentrated under reduced pressure. The residue was purified bysilica gel column chromatography, eluted with PE/EA (5:1) to afford thedesired compound (360 mg, 83%) as a white solid. ESI-MS m/z: 293.00[M+H]⁺.

Intermediate 39 Step f

To a solution of the compound from step e (360 mg, 1.2 mmol) in EtOH (4mL) and DMF (4 mL) was added Pd(OAc)₂ (83 mg, 0.4 mmol), DPPP (304 mg,0.74 mmol) and TEA (1 mL) in a pressure tank. The mixture waspressurized to 10 atm with carbon monoxide at 100° C. overnight. Thereaction mixture was cooled to room temperature and filtered to removeinsoluble solids. The residue was purified by reverse flashchromatography to give the desired compound (300 mg, 85%) as whitesolid. ESI-MS m/z: 287.10 [M+H]⁺.

Intermediate 39 Step g

Into a 50 mL round-bottom flask were added the compound from step f (298mg, 1 mmol), LiOH (125 mg, 5.2 mmol), MeOH (5 mL) and H₂O (1 mL) at roomtemperature. The resulting mixture was stirred for 2h at roomtemperature under nitrogen atmosphere. The reaction was monitored byLCMS. The mixture was acidified to pH 6 with HCl (aq.). The resultingmixture was concentrated under reduced pressure. The residue waspurified by reverse flash chromatography to give the desired compound(245 mg, 91%) as off-white solid. ESI-MS m/z: 259.10 [M+H]⁺.

Intermediate 40

Intermediate 40 Steps a and b

A mixture of methyl 4-amino-3-methoxybenzoate (50.00 g, 275.95 mmol) andacrolein (30.94 g, 551.87 mmol) in HCl (100 mL) and AcOH (100 mL) wasstirred for 1 h at 100° C. under nitrogen atmosphere. The resultingmixture was concentrated under vacuum used in next step directly. ESI-MSm/z: 204.10 [M+H]⁺.

A mixture of the compound from step a, ethyl iodide (76.76 g, 492.16mmol) and Cs₂CO₃ (240.52 g, 738.20 mmol) in DMF was stirred overnight atroom temperature. The resulting mixture was filtered, the filter cakewas washed with ethyl acetate (3×300 mL). The filtrate was concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography, eluted with hexane/EA (1:1) to afford the crude product(25 g) as a yellow solid. ESI-MS m/z: 232.05 [M+H]⁺.

Intermediate 40 Step c

A mixture of ethyl 8-methoxyquinoline-6-carboxylate (14.00 g, 60.54mmol) and NIS (54.48 g, 242.16 mmol) in AcOH (300 mL) was stirred for 16h at 100° C. under nitrogen atmosphere. The aqueous layer was extractedwith EA. The resulting mixture was concentrated under vacuum. Theresidue was purified by silica gel column chromatography and reverseflash chromatography to afford the desired product (5.5 g, 14%) as ayellow solid. ESI-MS m/z: 358.00 [M+H]⁺.

Intermediate 40 Step d

A mixture of the compound from step c (4.30 g, 12.04 mmol) and(1,10-Phenanthroline)(trifluoromethyl)copper(I) (5.65 g, 18.06 mmol) inDMF (70 mL) was stirred for 4 hours at 110° C. under nitrogenatmosphere. The mixture was allowed to cool down to room temperature.The resulting mixture was poured into water, extracted with EA. Thecombined organic layers were washed with brine and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with PE/EA (4:1) to afford the desired product(2.6 g, 72%) as a light yellow solid. ESI-MS m/z: 300.15 [M+H]⁺.

Intermediate 40 Step e

A mixture of the compound from step d (2.60 g, 8.69 mmol) and LiOH (2.08g, 86.85 mmol) in MeOH (30 mL) and H₂O (10 mL) was stirred for 2 hoursat room temperature. The mixture was acidified to pH 4 with HCl (2 Maq.). The precipitated solids were collected by filtration and washedwith water to afford the desired product (2.29 g, 97%) as a white solid.ESI-MS m/z: 271.95 [M+H]⁺.

Intermediate 41

The following compound was prepared in an analogous fashion toIntermediate 39. The residue was purified by reverse flashchromatography to afford the desired product (40 mg, 89%) as a lightyellow solid. ESI-MS m/z: 247.10 [M+H]⁺.

Intermediate 42

The following compound was prepared in an analogous fashion toIntermediate 10. The crude residue was purified by reverse flashchromatography to afford the desired product (130 mg, 72%) as a whitesolid. ESI-MS m/z: 247.10 [M+H]⁺.

Intermediate 43

Intermediate 43 Steps a and b

To a 40 mL vial equipped with a stir bar was added methyl3-methoxyquinoline-6-carboxylate (400 mg, 1.841 mmol) and the materialwas dissolved in DCM. mCPBA (883 mg, 3.68 mmol) was added, and thereaction was stirred overnight for 20 hr. The reaction was diluted withDCM and quenched with sodium thiosulfate. DCM/MeOH extraction with aphase separator cartridge, and the organics were concentrated. Thereside was purified by automated column chromatography (silica gel,0-20% methanol in DCM) to afford the desired compound (361 mg, 84%).ESI-MS m/z: 234.08 [M+H]⁺.

To a 50 mL rbf containing a stir bar was step a (143 mg, 0.613 mmol),and the solid was dissolved in DCM. POCl₃ (114 μl, 1.226 mmol) wasadded, the flask equipped with a condenser, and the reaction heated to45° C. for 2 hr. The reaction was diluted with DCM and quenched withwater and saturated sodium bicarbonate. DCM and DCM/MeOH extraction witha phase separator cartridge, and the organics were concentrated. Thereside was purified by automated column chromatography (silica gel,0-90% EtOAc in hexanes) to afford the desired compound (102 mg, 66%).ESI-MS m/z: 252.04 [M+H]⁺.

Intermediate 43 Step c

To a 20 mL vial equipped with a stir bar was added Pd(PPh₃)₄ (4.59 mg,3.97 μmol), step b (40 mg, 0.159 mmol), potassium carbonate (65.9 mg,0.477 mmol) and methylboronic acid (47.6 mg, 0.795 mmol). The vial waspurged with N₂ for 10 minutes. 1,4-Dioxane (1.060 ml) was added and thereaction was heated to 100° C. for 18 hrs. The reaction was diluted withEtOAc and quenched with water. EtOAc and DCM/MeOH extraction with aphase separator cartridge, and the organics were concentrated. Thereside was purified by automated column chromatography (silica gel,0-80% EtOAc in hexanes) to afford the desired compound (36 mg, 98%).ESI-MS m/z: 232.10 [M+H]⁺.

Intermediate 43 Step d

To a 20 mL vial equipped with a stir bar was added step c (36 mg, 0.156mmol), and the material was dissolved in THF:MeOH:Water (1:1:1, 0.2 M).Lithium hydroxide hydrate (32.7 mg, 0.778 mmol) was added, and thereaction was warmed to 45° C. and monitored by LCMS. The vial was cooledto 0° C. and acidified to pH=4 with 2 M HCl. No precipitate,concentrated organics in vacuo. Solid precipitated. Cooled to 0° C.,further precipitation. Solid collected via vacuum filtration and driedon high vacuum to afford the desired pdt as a white solid (18 mg, 53%).ESI-MS m/z: 217.82 [M+H]⁺.

Example 233

(S)-7-(4-fluorophenyl)-3-(hydroxymethyl)-5-((S)-1,1,1-trifluoro-2-hydroxy-3-(8-methoxy-2,3-dimethylquinoline-6-carboxamido)propan-2-yl)-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide,hydrochloride salt (20 mg, 0.032 mmol) was dissolved in DCM (1 ml) andDAST (1 eq, 4.2 μl, 0.032 mmol) was added at 0° C. The reaction mixturewas allowed to stir at 0° C. for 2 h. The reaction mixture was quenchedby adding MeOH and the solution was concentrated and purified by HPLC togive(S)-3-(chloromethyl)-7-(4-fluorophenyl)-5-((S)-1,1,1-trifluoro-2-hydroxy-3-(8-methoxy-2,3-dimethylquinoline-6-carboxamido)propan-2-yl)-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide(3.68 mg, 5.69 μmol, 17.88% yield). ESI-MS m/z: 647.074 [M+H]⁺.

Intermediate 44

The above example was prepared in an analogous sequence to Intermediate36 as a mixture of diastereomers to afford the desired amino alcohol(320 mg). ESI-MS m/z: 426.04 [M+H]⁺.

Intermediate 45

Intermediate 45 Step a

Into a 250 mL round-bottom flask were added methyl7-bromo-5-iodo-2,3-dihydrofuro[2,3-c]pyridine-3-carboxylate (586 mg,1.526 mmol), tetrahydrofuran (85 mL) and ((chloromethoxy)methyl)benzene(6.94 ml, 50.6 mmol). The mixture was cooled to −78° C. and lithiumdiisopropylamide (2 M solution in tetrahydrofuran/heptane/ethylbenzene,9.27 mL, 18.54 mmol) was added dropwise over 15 min. The mixture wasallowed to stir at −78° C. for 1 h, and was then warmed to 23° C. for 30min. The reaction was diluted with EtOAc (100 mL) and water (50 mL). Thelayers were separated and the aqueous layer was extracted with EtOAc(3×50 mL), the combined organics were dried over Na₂SO₄, filtered andconcentrated. The crude material was purified by automated columnchromatography (silica gel, 0-10% EtOAc in hexanes) to afford the titlecompound (4.69 g, 55%). ESI-MS m/z: 503.55/505.47 [M+H]⁺.

Intermediate 45 Step b

Into a 100 mL round-bottom flask were added the compound from step a(4.69 g, 9.30 mmol), and chloroform (47 mL). The mixture was cooled to0° C. and methanesulfonic acid was added. The mixture was allowed towarm to room temperature with stirring for 2 h and monitored by LCMS.The reaction was diluted with EtOAc and water. The aqueous wasextracted, the combined organics were washed with NaHCO₃ dried overNa₂SO₄, filtered and concentrated. The material was purified byautomated column chromatography (silica gel, 0-20% EtOAc in hexanes) toafford the title compound (2.25 g, 58%). ESI-MS m/z: 413.46/415.42[M+H]⁺.

Intermediate 45 Step c

Into a 50 mL round-bottom flask were added the compound from step b(2.12 g, 5.12 mmol), followed by dichloromethane (21 mL) then2,6-lutidine (890 μL, 7.68 mmol). The mixture was cooled to −78° C.,then triflic anhydride (1 M solution in dichloromethane, 5.63 mL 5.63mmoL) was added dropwise over 10 min. The mixture was allowed to stir at−78° C. for 30 min and monitored by LCMS. The mixture was warmed to 0°C. and was then diluted with water and dichloromethane. The layers wereseparated and the aqueous layer was washed with dichloromethane (3×25mL), the combined organics were washed with NaHCO₃ (sat. aq.) (2×mL)then concentrated. The resulting oil was dissolved in tetrahydrofuran (2mL), and cooled to 0° C. To this mixture tetrabutylammonium fluoride (1M in tetrahydrofuran, 5.37 mL, 5.37 mmol) was added, and the mixtureallowed to warm to room temperature for 19 h. The reaction was dilutedwith EtOAc and water. The aqueous was extracted with EtOAc (3×20 mL),the combined organics were dried over Na₂SO₄ and concentrated. Thematerial was purified by automated column chromatography (silica gel,0-10% EtOAc in hexanes) to afford the title compound (2.00 g, 94%).ESI-MS m/z: 415.47/417.40 [M+H]⁺.

Intermediate 45 Step d

Into a 100 mL round-bottom flask containing the compound from step c(2.00 g, 4.81 mmol), was added ammonia (7 M solution in MeOH, 20.6 mL,144 mmol). The mixture was heated to 45° C. for 1.5 h and monitored byLCMS. The mixture was cooled to room temperature, concentrated and theyellow solids were carried forward to the next step without furtherpurification (1.93 g, >99%). ESI-MS m/z: 400.46/402.41 [M+H]⁺.

Intermediate 45

The above compound was prepared in an analogous sequence to Intermediate36 as a mixture of diastereomers to afford the desired amino alcohol(1.056 g, 85%). ESI-MS m/z: 417.97 [M+H]⁺.

Intermediate 46

Intermediate 46 Step a

To a 250 mL round bottom flask equipped with a stir bar was added methyl7-bromo-5-iodo-2,3-dihydrofuro[2,3-c]pyridine-3-carboxylate (2.044 g,5.32 mmol) and the material was dissolved in THE (21.29 ml). The flaskwas cooled it −78° C., and (bromodifluoromethyl)trimethylsilane (1.656ml, 10.65 mmol) followed by dropwise addition of lithiumdiisopropylamide (2.93 ml, 5.86 mmol). The reaction was stirred for 1 hrand −78° C., and then allowed to warm to room temperature. After 1.5 hrthe reaction was diluted with EtOAc and quenched with sat. ammoniumchloride. The aqueous was extracted with EtOAc, and the organics dried,filtered and concentrated. The crude material was purified by automatedcolumn chromatography (silica gel, 0-30% EtOAc in hexanes, large columnvolume) to afford the desired product (295.6 mg, 13%). ESI-MS m/z:433.87/435.86 [M+H]⁺.

Intermediate 46 Step b

To a 40 mL vial containing step a (393.9 mg, 0.908 mmol) was added astir bar and ammonia (3890 μl, 27.2 mmol) was added. The reaction washeated to 45° C. and monitored by LCMS (1.5 hr complete). The stir barwas removed, and the reaction concentrated. The material was tritteredwith DCM to afford a light brown solid as the desired product (360 mg,96%). ESI-MS m/z: 418.87/420.87 [M+H]⁺.

Intermediate 46

The following compound was prepared in an analgous fashion toIntermediate 36 to afford the desired amino alcohol as a mixture ofdiastereomers (105 mg, 92%). ESI-MS m/z: 436.11 [M+H]⁺.

The following Table 2 contains examples that were prepared with thesimilar method to Example 1 Step c (PyBOP or HATU). The amine couplingpartners were prepared according to Intermediates 44-46 or by analogousprocedures as a mixture of diastereomers. The majority of compounds werepurified by Gilson prep-HPLC. If depicted as a single diasteromer, thediastereomers were separated on Gilson prep-HPLC. The aryl acid couplingpartners were prepared according to Intermediates 1-46 or by analogousprocedures with slight modifications, and also prepared according toprocedures found in U.S. patent application Ser. No. 16/930,622.

TABLE 2 MS⁺ Example Structure m/z 234

638.21 235

625.21 236

628.22 237

599.30 238

647.36 239

667.37 240

608.13 241

604.31 242

648.21 243

638.21 244

625.19 245

628.22 246

599.29 247

647.36 248

667.37 249

608.12 250

604.30 251

648.21 252

657.19 253

650.18 254

621.16 255

660.15 256

669.13 257

689.14 258

630.10 259

650.13 260

669.92 261

625.85 262

619.97 263

619.97 264

679.07 265

679.07 266

650.01 267

650.01 268

611.06 269

611.06 270

637.06 271

637.06 272

640.15 273

640.15 274

659.11 275

659.11 276

616.02 277

616.02 278

660.08 279

660.08 280

640.08 281

640.08 282

602.82 283

603.11 284

650.96 285

651.35 286

670.94 287

670.94 288

641.82 289

611.89 290

612.04 291

632.15 292

632.10 293

651.98 294

651.93 295

631.91 296

629.03 297

629.13 298

607.86 299

607.91

The following Table 3 contains examples that were prepared with thesimilar method to Example 1 Step c (PyBOP or HATU). The majority ofcompounds were purified by Gilson prep-HPLC, and some were purified byautomated column chromatography (silica gel). The aryl acid couplingpartners were prepared according to Intermediates 1-43, or by analogousprocedures with slight modifications and also prepared according toprocedures found in U.S. patent application Ser. No. 16/930,622.

TABLE 3 MS⁺ Example Structure m/z 300

658.34 301

648.35 302

586.24 303

596.20 304

601.94 305

627.83 306

643.81 307

609.83 308

611.84 309

600.89 310

684.95 311

667.91 312

586.14 313

566.03 314

669.14 315

671.15 316

561.2 317

605.30 318

560.10 319

605.03 320

586.20 321

640.20 322

628.40 323

628.40 324

662.40 325

619.15 326

599.22 327

635.17 328

681.02 329

644.35 330

644 .34 331

612.26 332

556.23

The following Table 4 contains examples that were prepared with thesimilar method to Example 1 step c (PyBOP or HATU). The majority ofcompounds were purified by Gilson prep-HPLC, and some were purified byautomated column chromatography (silica gel). The aryl acid couplingpartners were commercially available.

TABLE 4 MS⁺ Example Structure m/z 333

578.00 334

519.0 335

562.00 336

587.05 337

509.05 338

610.85 339

600.57 340

594.05 341

592.90 342

535.05 343

543.85 344

519.00 345

576.85 346

559.80 347

611.85 348

560.85 349

492.85 350

576.80 351

544.90 352

540.90 353

561.90 354

586.90 355

507.15 356

508.14 357

559.1 358

558.16 359

561.05 360

544.10 361

572.15 362

507.05 363

494.00 364

495.00 365

545.10 366

543.10 367

562.10 368

560.05 369

544.05 370

544.05 371

561.05 372

562.10 373

557.10 374

545.05 375

559.16 376

560.10 377

561.05 378

544.05 379

577.05 380

544.05 381

509.05 382

548.05 383

544.01 384

544.05 385

557.10 386

524.05 387

561.15 388

544.05 389

510.95 390

520.05 391

493.00 392

544.10 393

545.10 394

591.05 395

574.10 396

510.10 397

561.10 398

557.20 399

554.20 400

560.10 401

556.20 402

543.20 403

544.10 404

493.10 405

561.10 406

505.10 407

506.20 408

554.10 409

544.10 410

519.20 411

507.20 412

552.10 413

543.20 414

577.10 415

542.20 416

520.05 417

545.05 418

544.05 419

594.00 420

544.05 421

505.05 422

561.05 423

548.05 424

558.15 425

507.05 426

494.05 427

552.05 428

535.05 429

544.10 430

574.10 431

525.05 432

569.10 433

545.10 434

578.00 435

574.00 436

545.00 437

553.10 438

561.05 439

545.05 440

614.20 441

578.18 442

552.17 443

646.38 444

570.29 445

584.01 446

569.96 447

675.98 448

600.32 449

600.31 450

600.18 451

588.16 452

584.19 453

598.20 454

587.89 455

637.85 456

637.92 457

637.85 458

584.19 459

535.05 460

558.10 461

534.20 462

565.17 463

628.33 464

628.34 465

592.02 466

544.20 467

558.08 468

558.10 469

558.10 470

577.92 471

574.03 472

561.90 473

522.20 474

508.17 475

601.17 476

539.15 477

551.16 478

587.15 479

601.92 480

617.90 481

548.08 482

587.14 483

601.15 484

605.12 485

619.14 486

556.49 487

592.02 488

544.20 489

545.37 490

576.34

Intermediate 47

Intermediate 47 Step a

A mixture of(R)-7-bromo-5-iodo-3-methyl-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide(3.5 g, 9.14 mmol), cyclopropyl vinyl boronic acid pinacol ester (2.13g, 10.96 mmol), Pd(dppf)Cl₂ (1.34 g, 1.83 mmol), K₂CO₃ (3.16 g, 22.84mmol) in dioxane (18 mL) and H₂O (2 mL) was stirred for 2 hours at 90°C. under nitrogen atmosphere. The reaction was monitored by LCMS. Theresulting mixture was filtered, the filter cake was washed with EA. Thefiltrate was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (eluted 50% ethyl acetatein hexanes) to afford the desired product (2.2 g, 74%) as a brown oil.ESI-MS m/z: 322.90 [M+H]⁺.

Intermediate 47 Step b

A mixture of the compound from step a (1.5 g, 4.64 mmol),4-fluorophenylboronic acid (0.97 g, 6.96 mmol), Pd(PPh₃)₂Cl₂ (0.33 g,0.46 mmol), and Na₂CO₃ (1.23 g, 11.60 mmol) in THE (10 mL) and H₂O (2.5mL) was stirred for 1 h at 70° C. under nitrogen atmosphere. Thereaction was monitored by TLC. The resulting mixture was filtered, thefilter cake was washed with EA. The filtrate was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (eluted with 33% ethyl acetate in hexanes) to afford thedesired product (1.2 g, 76%) as a yellow oil. ESI-MS m/z: 339.05 [M+H]⁺.

Intermediate 47 Step c

A solution of the compound from step b (1.2 g, 3.54 mmol) in t-BuOH (40mL) and H₂O (40 mL) was cooled to 0° C. Methanesulfonamide (0.34 g, 3.54mmol) and AD-mix-β (8.29 g, 10.64 mmol) were added and the reaction wasstirred overnight at room temperature and monitored by LCMS. Theresulting mixture was poured into water and extracted with ethylacetate. The combined organic layers were concentrated under reducedpressure. The residue was purified by reverse flash chromatography (MeCNin water, 0.1% FA) to afford the desired product (1.1 g, 83%) as anoff-white solid. ESI-MS m/z: 373.25[M+H]⁺.

Intermediate 47 Step d

A solution of the compound from step c (1.7 g, 4.56 mmol) in DCM (70 mL)was cooled to 0 C°. TsCl (1.31 g, 6.84 mmol), TEA (1.39 g, 13.69 mmol)and DMAP (0.22 g, 1.82 mmol) were added and the reaction stirred for 3hours at room temperature. The reaction was monitored by LCMS. Themixture was acidified to pH 4 with HCl (2 M aq.). The resulting mixturewas extracted with CH₂C₁₂. The combined organic layers were concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (eluted with 66% ethyl acetate in hexanes) to afford thedesired product (1.6 g, 66%) as a yellow solid. ESI-MS m/z: 527.30[M+H]⁺.

Intermediate 47 Step e

Into a 250 mL round-bottom flask containing step d (1.6 g, 3.04 mmol)were added NH₃ (68 mL, 156 eq, 7N in MeOH) at room temperature. Theresulting mixture was stirred for 20 hr at room temperature andmonitored by LCMS. The solvent was removed, and the crude mixture wasdissolved in EtOAc. The organics were washed 3× with sat. sodiumbicarbonate, and the organics were concentrated. The crude material wastriturated with DCM to afford the desired product (590.9 mg, 52%) as anoff-white solid. ESI-MS m/z: 372.15 [M+H]⁺.

Intermediate 48

Intermediate 48 Step a

In a vial, the compound from Intermediate 36 Step b (6.5 g, 17 mmol) and1-fluoro-N-methoxy-N-methylcyclopropane-1-carboxamide (3 g, 20.39 mmol)were dissolved in THE (121 ml) and cooled to 0° C. Isopropylmagnesiumchloride (16.99 ml, 34.0 mmol) was added slowly and the reaction wasallowed to stir at 0° C. After stirring for 3 hours, water added at 0°C. and reaction allowed to warm to RT. The aqueous layer washed withEtOAc and combined organic layer dried over MgSO₄ and concentrated. Thecrude reaction mixture purified by silica gel column chromatographyeluting with 0-50% EtOAc/Hexanes to give the title compound (2.8 g, 48%)as a foaming white solid. ESI-MS m/z: 343.17 [M+H]⁺.

Intermediate 48 Step b

In a vial, the compound from step a (1.4 g, 4.08 mmol),(4-fluorophenyl)boronic acid (0.685 g, 4.90 mmol), PdCl₂(dppf) DCM(0.200 g, 0.245 mmol), and K₂CO₃ (1.692 g, 12.24 mmol) were dissolved in1,4-Dioxane (16.32 ml) and Water (4.08 ml). The reaction was spargedwith N₂ and sealed. The reaction was heated to 90° C. After 4 hours, thereaction was cooled to RT and water was added. The aqueous layer waswashed with EtOAc. The combined organic layer was washed with water andbrine before drying over MgSO₄ and concentrating. The crude reactionmixture purified by silica gel column chromatography eluting with 0-70%EtOAc/Hexanes to give the title compound (1.12 g, 77%). ESI-MS m/z:359.06 [M+H]⁺.

Intermediate 48 Step c

A vial was charged with methyltriphenylphosphonium bromide (3.00 g, 8.41mmol), and THE (18.7 ml), then cooled to 0° C. in an ice bath. Potassiumtert-butoxide (0.906 g, 8.07 mmol) was added slowly as a solution in THE(2 mL), and the now yellow suspension was stirred for 30 min at 0° C.The compound from step b (1.205 g, 3.36 mmol) was added as a solution inTHE (15 ml). The reaction was allowed to stir at 0° C. for 2 hoursbefore allowing to warm to room temperature and stir an additional 2hours. The reaction was quenched upon addition of MeOH (5 mL) then Water(20 mL). The solution was diluted with diluted with EtOAc and the layerswere separated. The aqueous layer washed with EtOAc and the combinedorganic layer was washed with brine, dried over MgSO₄, and concentrated.The crude reaction mixture purified by silica gel column chromatographyeluting with 0-80% EtOAc/Hexanes to give the title compound (741 mg,61.8%). ESI-MS m/z: 357.39 [M+H]⁺.

Intermediate 48 Step d

The above compound was prepared in an analogous fashion to Intermediate36 Step e. The reaction mixture was purified by silica gel columnchromatography eluting with 0-80% EtOAc/Hex to afford the title compound(480 mg, 59%). ESI-MS m/z: 391.33 [M+H]⁺.

Intermediate 48 Step e

The above compound was prepared in an analogous fashion to Intermediate36 Step f. After aqueous workup, the title compound (585 mg, 87%) wastaken forward without further purification. ESI-MS m/z: 545.38 [M+H]⁺.

Intermediate 48 Step f

The above compound was prepared in an analogous fashion to Intermediate36 Step g. After aqueous workup, the title compound (381 mg, 91%) wastaken forward without further purification. ESI-MS m/z: 390.35 [M+H]⁺.

Intermediate 49

The above compound was prepared in an analogous fashion to Intermediate47 to afford the desired amino alcohol (473 mg, 64%). ESI-MS m/z: 434.40[M+H]⁺.

Intermediate 50

The above compound was prepared in an analogous fashion to Intermediate36 to afford the desired amino alcohol (493 mg, 92%). ESI-MS m/z: 452.25[M+H]⁺.

Intermediate 51

The above compound was prepared in an analogous fashion to Intermediate47 to afford the desired amino alcohol (246 mg, 83%). ESI-MS m/z: 424.31[M+H]⁺.

Intermediate 52

The above compound was prepared in an analogous fashion to Intermediate47 to afford the desired amino alcohol (147 mg, 95%). ESI-MS m/z: 422.36[M+H]⁺.

Intermediate 53

Intermediate 53 Step a

Into a 500 mL round-bottom flask were added2-chloro-5-fluoropyridin-3-ol (10 g, 68 mmol), K₂CO₃ (19 g, 136 mmol),12 (19 g, 75 mmol) and H₂O (200 mL) at room temperature. The resultingmixture was stirred for overnight at room temperature under nitrogenatmosphere and monitored by LCMS. The reaction was quenched by theaddition of sat. sodium hyposulfite (aq.) at room temperature andextracted with EtOAc. The residue was purified by silica gel columnchromatography (eluted with 9% ethyl acetate in hexanes) to afford thedesired compound (16.7 g, 90%) as a white solid. ESI-MS m/z: 273.90[M+H]⁺.

Intermediate 53 Step b

Into a 250 mL vial were added the compound from step a (12.4 g, 45mmol), (2-methyloxiran-2-yl)methyl 4-methylbenzenesulfonate, K₂CO₃ (13g, 91 mmol), KI (9 g, 54 mmol) and DMF (50 mL) at room temperature. Theresulting mixture was stirred for overnight at 50° C. under nitrogenatmosphere. The reaction was monitored by LCMS. The reaction wasquenched with water and the aqueous layer was extracted with EtOAc. Theresulting mixture was concentrated under reduced pressure. The residuewas purified by reverse flash to afford the desired compound (9.6 g,63%) as white solid. ESI-MS m/z: 343.80 [M+H]⁺.

Intermediate 53 Step c

Into a 50 mL 3-necked round-bottom flask were added the compound fromstep b (1.5 g, 4 mmol), THE (15 mL) and LDA (2.4 mL, 5 mmol) at 0° C.The resulting mixture was stirred for 1h at room temperature undernitrogen atmosphere. The reaction was monitored by TLC. The reaction wasquenched by the addition of sat. NH₄Cl (aq.) at 0° C. The aqueous layerwas extracted with EtOAc. The residue was purified by reverse flashafford the desired compound (430 mg, 29%) as white solid. ESI-MS m/z:343.80 [M+H]⁺.

Intermediate 53 Step d

Into a 100 mL round-bottom flask were added the compound from step c(400 mg, 1 mmol), acetone (10 mL) and Jones Reagent (1 mL) at 0° C. Theresulting mixture was stirred for 3 hr at room temperature undernitrogen atmosphere. The reaction was monitored by LCMS. The aqueouslayer was extracted with EtOAc and concentrated under reduced pressure.The crude product was used in the next step directly without furtherpurification. ESI-MS m/z: 357.85 [M+H]⁺.

Intermediate 53 Step e

Into a 50 mL round-bottom flask were added the crude compound from stepd, CDI (726 mg, 4 mmol) and THE (5 mL) at room temperature. Theresulting mixture was stirred for 2 hr at room temperature undernitrogen atmosphere. Then the resulting mixture was added to NH₃.H₂O (50mL) dropwise and stirred for 1 hr. The reaction was monitored by TLC.The aqueous layer was extracted with EtOAc and the residue was purifiedby reverse flash to afford the desired compound (218.8 mg, 69%) as whitesolid. ESI-MS m/z: 356.90 [M+H]⁺.

Intermediate 54

The following compound was prepared in an analogous sequence toIntermediate 48 steps a and b. ESI-MS m/z: 336.95 [M+H]⁺.

Intermediate 55

The above compound was prepared in an analogous fashion to Intermediate36. After aqueous workup, the title compound (45 mg, 63%) was isolatedas a white solid. ESI-MS m/z: 386.40 [M+H]⁺.

Intermediate 56

The following compound was prepared in an analogous sequence toIntermediate 48 steps a and b steps a and b. ESI-MS m/z: 351.05 [M+H]⁺.

Intermediate 57

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by automated column chromatography (silica gel, 10% MeOH inDCM) to afford the desired product (1.80 g, 57%) as an off-white solid.ESI-MS m/z: 462.05 [M+H]⁺.

Intermediate 58

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 10% MeOH in DCM with NH₃) to affordthe desired product (508 mg, 74%) as a white solid. ESI-MS m/z: 468.05[M+H]⁺.

Intermediate 59

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 6% MeOH in DCM with NH₃) to afford thedesired product (450 mg, 72%) as a white solid. ESI-MS m/z: 434.10[M+H]⁺.

Intermediate 60

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 6% MeOH in DCM with NH₃) to afford thedesired product (468.9 mg, 92%) as a white solid. ESI-MS m/z: 433.95[M+H]⁺.

Intermediate 61

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 6% MeOH in DCM with NH₃) to afford thedesired product as a white solid. ESI-MS m/z: 434.10 [M+H]⁺.

Intermediate 62

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 6% MeOH in DCM with NH₃) to afford thedesired product as a white solid. ESI-MS m/z: 452.00 [M+H]⁺.

Intermediate 63

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid pinacol ester. The arylboronic acid pinacol ester was prepared from the corresponding bromideby Pd-catalyzed borylation. The crude material was purified by prep-TLC(silica gel, 6% MeOH in DCM with NH₃) to afford the desired product (355mg, 67%) as a white solid. ESI-MS m/z: 452.10 [M+H]⁺.

Intermediate 64

The above compound was prepared in an analogous fashion to Intermediate36 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 6% MeOH in DCM with NH₃) to afford thedesired product (570.3 mg, 87%) as a white solid. ESI-MS m/z: 452.10[M+H]⁺.

Intermediate 65

Into a 40 mL sealed tube were added 3-bromo-7-fluoro-1-benzothiophene(1.29 g, 5.60 mmol), bis(pinacolato)diboron (1.42 g, 5.59 mmol), KOAc(1.65 g, 16.81 mmol), Pd(dppf)Cl₂.CH₂Cl₂ (365 mg, 0.45 mmol) and dioxane(18 mL) at room temperature. The resulting mixture was stirred for 1 hrat 110° C. under nitrogen atmosphere. The reaction was monitored byLCMS. The mixture was allowed to cool down to room temperature.(R)-7-bromo-3-methyl-5-(3,3,3-trifluoroprop-1-en-2-yl)-2,3-dihydrofuro[2,3-c]pyridine-3-carboxamide(787 mg, 2.24 mmol), K₂CO₃ (0.93 g, 6.72 mmol), Pd(dppf)Cl₂.CH₂C₁₂ (365mg, 0.45 mmol) and H₂O (2 mL) was added and stirred for another 2 hr at80° C. The resulting mixture was filtered, the filter cake was washedwith EA. The filtrate was concentrated under reduced pressure. Theresidue was purified by reverse flash chromatography to afford thedesired product (460 mg, 48%) as a brown oil. ESI-MS m/z: 422.95 [M+H]⁺.

Intermediate 66

The above compound was prepared in an analogous fashion to Intermediate36 with Intermediate 65. The crude material was purified by prep-TLC(silica gel, 8% MeOH in DCM with NH₃) to afford the desired product(251.3 mg, 73%) as a white solid. ESI-MS m/z: 456.00 [M+H]⁺.

Intermediate 67

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 10% MeOH in DCM with NH₃) to affordthe desired product (291.1 mg, 61%) as a white solid. ESI-MS m/z: 440.00[M+H]⁺.

Intermediate 68

The above compound was prepared in an analogous fashion to Intermediate47 and Intermediate 65 with the corresponding aryl boronic acid. Thecrude material was purified by prep-TLC (silica gel, 6% MeOH in DCM withNH₃) to afford the desired product (114.2 mg, 49%) as a white solid.ESI-MS m/z: 428.05 [M+H]⁺.

Intermediate 69

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, MeOH in DCM with NH₃) to afford thedesired product (220 mg, 64%) as a white solid. ESI-MS m/z: 406.10[M+H]⁺.

Intermediate 70

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, MeOH in DCM with NH₃) to afford thedesired product (267.8 mg, 64%) as a white solid. ESI-MS m/z: 406.10[M+H]⁺.

Intermediate 71

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, MeOH in DCM with NH₃) to afford thedesired product (262.1 mg, 53%) as a white solid. ESI-MS m/z: 406.15[M+H]⁺.

Intermediate 72

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, 6% MeOH in DCM with NH₃) to afford thedesired product (250 mg, 68%) as a white solid. ESI-MS m/z: 424.00[M+H]⁺.

Intermediate 73 Steps a and b

To a vial were added methyl 3-amino-4-fluorobenzoate (50 mg, 0.296mmol), isocyanatocyclopropane (24.56 mg, 0.296 mmol) and EtOH (2 mL).The resulting mixture was stirred at rt for 20 hr. After removed thesolvent, the residue was purified by automated column chromatography(silica gel 4 g column, eluting with 0-50% EtOAc/hexanes) to obtain thedesired product (30 mg, 40.2%) as a pale-yellow foam. ESI-MS m/z: 253.10[M+H]⁺. To a vial with solvents of THF/H₂O (2 mL, 9:1) were added step a(90 mg, 0.357 mmol) and 2N NaOH (1 mL). After stirred at rt overnight,the mixture was neutralized to pH˜3 by adding 1N HCl. The solid wasprecipitated out. After filtered and dried in oven, the desired product(85 mg, 100%) was obtained. ESI-MS m/z: 239.20 [M+H]⁺.

Intermediate 74

A 2 mL MW vial was charged with ethyl6-hydroxyimidazo[1,2-a]pyridine-2-carboxylate (60 mg, 0.291 mmol) and2-bromopropane (165 μl, 1.164 mmol) and DMF (0.582 ml). Cesium carbonate(284 mg, 0.873 mmol) was added subsequently. The reaction vial wascapped and heated at 140° C. for 2 h in a microwave reactor. Thereaction mixture was diluted with ethyl acetate and water. The organiclayer was washed with brine and dried and concentrated and used in thenext step without further purification. ESI-MS m/z: 249.12 [M+H]⁺.

Intermediate 75

Ethyl 3-bromo-2-oxopropanoate (68.5 μl, 0.546 mmol) and5-(trifluoromethoxy)pyridin-2-amine (81 mg, 0.455 mmol) were dissolvedin DME (1.137 ml) and MeOH (1.137 ml). The reaction mixture was heatedat 80° C. for 18h. The solvent was removed, and the residue was purifiedby flash chromatography to give the desired product (78 mg, 62.6%).ESI-MS m/z: 275.057 [M+H]⁺.

Intermediate 76

Methyl 2-(1-hydroxycyclopropyl)-7-methoxy-2H-indazole-5-carboxylate (55mg, 0.210 mmol) was dissolved in dichloromethane (1.049 ml) at 0° C.Thionyl chloride (61.2 μl, 0.839 mmol) was added and the mixture wasallowed to stir at 50° C. for 18 h. The reaction mixture was purified byflash chromatography directly to give the desired product (34 mg,57.8%). ESI-MS m/z: 281.061 [M+H]⁺.

Intermediate 77

Intermediate 77 Step a

Into a 100 mL round-bottom flask were added5-bromo-1-methoxy-2-methyl-3-nitrobenzene (1 g, 4 mmol), NBS (0.8 g, 4.5mmol), AIBN (0.13 g, 0.8 mmol) and CCl₄ (20 mL) at room temperature. Theresulting mixture was stirred for overnight at 80° C. under nitrogenatmosphere and monitored by LCMS. The reaction was quenched by theaddition of water and the resulting mixture was extracted with EtOAc.The combined organics were dried over sodium sulfate, filtered andconcentrated. The crude product was used in the next step directlywithout further purification. ESI-MS m/z: 325.90 [M+H]⁺.

Intermediate 77 Step b

Into a 100 mL round-bottom flask were the compound from step a (1.33 g,4 mmol,), aminocyclopropane (2.34 g, 41 mmol), K₂CO₃ (1.13 g, 8 mmol)and DMF (10 mL) at room temperature. The resulting mixture was stirredfor 3h at room temperature under nitrogen atmosphere. The reaction wasmonitored by LCMS. The aqueous layer was extracted with EtOAc. Theresidue was purified by silica gel column chromatography (eluted with16% ethyl acetate in hexanes) to afford the desired compound (1.07 g,87%) as a white solid. ESI-MS m/z: 301.00 [M+H]⁺.

Intermediate 77 Step c

Into a 100 mL 3-necked round-bottom flask were added the compound fromstep b (500 mg, 1.7 mmol) and THE (6 mL) at 0° C. NaOH (664 mg, 17 mmol)in H₂O (6 mL) was added over 2 min and then Zinc (326 mg, 5 mmol) wasadded in 6 portions during 3 hr and stirred for another 1 hr. Thereaction was monitored by LCMS. The mixture was acidified to pH 3 withHCl (aq.). The aqueous layer was extracted with CH₂Cl₂, dried overanhydrous Na₂SO₄. After filtration, the filtrate was concentrated underreduced pressure and washed with hexane and resulted in the desiredcompound (310 mg, 70%) as white solid. ESI-MS m/z: 267.00 [M+H]⁺.

Intermediate 77 Step d

Into a 30 mL pressure tank reactor were added the compound from step c(240 mg, 0.9 mmol), dppp (222 mg, 0.5 mmol), Pd(OAc)₂ (60 mg, 0.3 mmol),DMF (4 mL), EtOH (4 mL) and TEA (1 mL) at room temperature. Theresulting mixture was stirred for overnight at 100° C. under COatmosphere at 15 atm. The reaction was monitored by LCMS. The resultingmixture was filtered, the filter cake was washed with EA. The filtratewas concentrated under reduced pressure. The residue was purified byreverse phase silica gel chromatography (MeCN/H₂O) to afford the desiredcompound (210 mg, 90%) as white solid. ESI-MS m/z: 261.10 [M+H]⁺.

Intermediate 77 Step e

Into a 50 mL round-bottom flask were added the compound from step d (470mg, 1.8 mmol), lithium hydroxide (216 mg, 9 mmol), methanol (10 mL) andH₂O (3 mL) at room temperature. The resulting mixture was stirred for 1hr at room temperature under nitrogen atmosphere. The reaction wasmonitored by LCMS. The mixture was acidified to pH 6 with HCl (aq.). Theresulting mixture was washed with water and evaporated to give thedesired compound (317.8 mg, 76%) as yellow solid. ESI-MS m/z: 233.10[M+H]⁺.

Intermediate 78

Intermediate 78 Step a

Into a 50 mL round-bottom flask were added methyl3-[(cyclopropylamino)methyl]-5-methoxy-4-nitrobenzoate (3 g, 11 mmol),KOH (6 g, 107 mmol), MeOH (20 mL) and H₂O (2 mL) at room temperature.The resulting mixture was stirred for 72 hr at 80° C. under nitrogenatmosphere. The reaction was monitored by TLC. The mixture was acidifiedto pH 6 with HCl (aq.). The residue was purified by reverse phase silicagel chromatography (MeCN/H₂O) to afford the desired compound (580 mg,21%) as white solid. ESI-MS m/z: 263.10 [M+H]⁺.

Intermediate 78 Step b

Into a 40 mL vial were added the compound from step a (1.28 g, 5 mmol)and AcOH (6 mL) at room temperature. The resulting mixture was stirredfor 3 days at 120° C. under nitrogen atmosphere. The reaction wasmonitored by LCMS. The resulting mixture was concentrated under reducedpressure. The residue was purified by reverse phase silica gelchromatography (MeCN/H₂O) to afford the desired compound (0.96 g, 79%)as brown solid. ESI-MS m/z: 249.05 [M+H]⁺.

Intermediate 79

Intermediate 79 Step a

A solution of 5-bromo-7-methoxy-1H-indazole (3.00 g, 13.21 mmol) in MeCN(50 mL) was treated with 1-(benzenesulfonyl) cyclopropan-1-ol (3.14 g,15.85 mmol) and TEA (1.60 g, 15.85 mmol) for overnight at roomtemperature. The reaction was monitored by LCMS. The resulting mixturewas extracted with EtOAc and concentrated. The residue was purified bysilica gel column chromatography (eluted with 50% ethyl acetate inhexanes) to afford the desired compound (2.40 g, 64%) as a yellow solid.ESI-MS m/z: 283.00 [M+H]⁺.

Intermediate 79 Step b

A solution of the compound from step a (2.43 g, 8.58 mmol) in DCM (20mL) was treated with BAST (5.69 g, 25.74 mmol) for 1.5 hours at 0° C. tor.t. The reaction was monitored by LCMS and quenched with water. Theaqueous layer was extracted with EtOAc and concentrated. The residue waspurified by silica gel column chromatography (eluted with 50% ethylacetate in hexanes) to afford the desired compound (1.2 g, 49%) as ayellow solid. ESI-MS m/z: 285.00 [M+H]⁺.

Intermediate 79 Steps c and d

The following compound was prepared according to the same procedures asIntermediate 77 steps d and e to afford the desired product (312.6 mg,58%) as a white solid. ESI-MS m/z: 250.95 [M+H]⁺.

Intermediate 80

The following compound was prepared according to the same procedures asIntermediate 79 to afford the desired product (170.3 mg, 76%) as a whitesolid. ESI-MS m/z: 276.95 [M+H]⁺.

Intermediate 81

Intermediate 81 Steps a and b

A mixture of 5-bromo-1-fluoro-3-methyl-2-nitrobenzene (5 g, 21.37 mmol),cyclopropanol (2.5 g, 42.73 mmol) and Cs₂CO₃ (21 g, 64 mmol) in DMF (20mL) was stirred for 2 hr at 50° C. under N₂ atmosphere. The resultingmixture was extracted with EtOAc (3×200 mL). The combined organic layerswere washed with water, concentrated under reduced pressure to affordthe crude product without further purification.

A solution/mixture of the compound from step a (5 g, 18.38 mmol), Fe(10.3 g, 183.76 mmol) and NH₄Cl (9.8 g, 183.76 mmol) in EtOH (40 mL) andH₂O (20 mL) was stirred for 2 hr at 80° C. The residue was purified bysilica gel column chromatography (ethyl acetate in hexanes) to affordthe desired product (3.6 g, 81%) as a yellow solid. ESI-MS m/z: 242.00[M+H]⁺.

Intermediate 81 Steps c and d

To the compound from step b (1.5 g, 6.2 mmol) was added to a 50% aqueoussolution of fluoroboric acid (9.8 mL) at room temperature and stirredfor 5 min. The mixture was cooled in an ice bath for 10 min and anaqueous solution of NaNO₂ (900 mg, 13.04 mmol) in 1.7 mL H₂O was addedto the mixture. The reaction mixture was stirred at 10° C. for 30 min,during which the product precipitated. The cooled reaction mixture wasfiltered through a Buchner funnel and the solid product washed withsmall amounts of H₂O, MeOH and Et₂O, and dried under high vacuum toobtain the crude product as a brown solid. ESI-MS m/z: 253.00 [M+H]⁺.

In a dry flask, 18-crown-6 (0.05 g, 0.2 mmol) and potassium acetate (2.9g, 29.4 mmol) were dried under high vacuum for 1 hr. CHCl₃ (70 mL) wasadded and stirred at room temperature for 10 min. Then step c was addedto the mixture in small portions under N₂ atmosphere. The reactionmixture was stirred at room temperature for 3 h, filtered and theresidue washed with CHCl₃. The filtrate was washed with water (3×40 mL)and the organic layer was dried over Na₂SO₄, filtered and concentratedin vacuo to give the crude product. The crude product was purified byflash column chromatography (hexane/EtOAc) to afford the desired product(1.8 g, 82%) as a yellow solid. ESI-MS m/z: 253.00 [M+H]⁺.

Intermediate 81 Steps e, f, g

The following intermediate was prepared in an analogous fashion toIntermediate 6 steps a-c to afford the desired product (330 mg, 87%) asa yellow solid. ESI-MS m/z: 269.10 [M+H]⁺.

Intermediate 82

Intermediate 82 Steps a-d

The following intermediate was prepared in an analogous fashion toIntermediate 81 steps a-d using cyclopropanemethanol to afford thedesired product (1.85 g, 66%) as a light-yellow solid. ESI-MS m/z:266.90 [M+H]⁺.

Intermediate 82 Step e-h

The following intermediate was prepared in an analogous fashion toIntermediate 79 steps a-d to afford the desired product (298 mg, 60%) asa white solid. ESI-MS m/z: 291.10 [M+H]⁺.

Intermediate 83

The following intermediate was prepared in an analogous fashion toIntermediate 6 to afford the desired product (260 mg, 69%) as a whitesolid. ESI-MS m/z: 247.00 [M+H]⁺.

Intermediate 84

The following intermediate was prepared in an analogous fashion toIntermediate 6 to afford the desired product. ESI-MS m/z: 231.00 [M+H]⁺.

Intermediate 85

Intermediate 85 Step a

A solution/mixture of methyl 6-chloro-5-methoxypyridine-3-carboxylate (1g, 4.96 mmol), ethynylcyclopropane (0.66 g, 9.92 mmol), Pd(PPh₃)₄ (1.15g, 0.99 mmol), CuI (0.47 g, 2.48 mmol,) and Et₃N (1.51 g, 14.88 mmol) in1,4-dioxane (30 mL) was stirred for overnight at 90° C. under N₂atmosphere. The residue was purified by silica gel column chromatography(eluted with 50% ethyl acetate in hexanes) to afford the desired product(1 g, 87%) as a yellow solid. ESI-MS m/z: 232.00 [M+H]⁺.

Intermediate 85 Steps b-d

To a solution of the compound from step a (1 g, 4.32 mmol) in CH₂Cl₂(100 mL) was added a solution of amino 2,4,6-trimethylbenzenesulfonate(1.21 g, 5.62 mmol) in CH₂Cl₂ (50 mL) under ice cooling, and thereaction mixture was stirred for another one hour. Et₂O (12 mL) wasadded to the reaction mixture to precipitate crystals. The filtrate wasfiltered off and then dried under reduced pressure to afford a crudeproduct as a pale-yellow solid. ESI-MS m/z: 247.00 [M+H]⁺. Asolution/mixture of the compound from step b (500 mg, 2.02 mmol) andK₂CO₃ (559 mg, 4.04 mmol) in MeOH (50 mL) was stirred for 2 hr at roomtemperature under N₂ atmosphere. The residue was purified by silica gelcolumn chromatography (eluted with ethyl acetate in hexanes) to affordthe desired product (220 mg, 44%) as a yellow solid. ESI-MS m/z: 247.00[M+H]⁺. A solution of the compound from step c (220 mg, 0.89 mmol) andLiOH (214 mg, 8.93 mmol) in MeOH (10 mL) and H₂O (10 mL) was stirred for2 hr at room temperature. The mixture was acidified to pH 5-6 with HCl.The resulting mixture was filtered, the filter cake was collected toafford the desired product (160 mg, 77%) as a white solid. ESI-MS m/z:233.00 [M+H]⁺.

Intermediate 86

Intermediate 86 Step a

A mixture of 3-methoxy-6-bromopyridin 2-ylamine (1.0 g, 3.6 mmol) andchloroacetaldehyde (1.2 mL, 50% wt in water) in EtOH (12 mL) was stirredat reflux for 1 hour, then concentrated under reduced pressure. Theresidue was partitioned between an saturated solution of sodiumbicarbonate and EtOAc. The organic layer was separated and washed withbrine, dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash chromatography (Si-PPC, MeOH/Et₂O, gradient 0:100 to1:99) to give the desired product as a beige solid (960 mg, 88%). ESI-MSm/z: 227.00 [M+H]⁺.

Intermediate 86 Steps b and c

The following example was prepared in an analogous fashion toIntermediate 77 to afford the desired product (340 mg, 49%) as a yellowsolid. ESI-MS m/z: 192.95 [M+H]⁺.

Intermediate 87

Intermediate 87 Steps a and b

To a solution of 2,6-dibromo-4-methoxypyridine (5 g, 18.73 mmol),tert-butyl carbamate (2.19 g, 18.73 mmol) and Cs₂CO₃ (12.21 g, 37.46mmol) in 1,4-dioxane (60 mL) was added Pd(OAc)₂ (0.42 g, 1.87 mmol) andXantphos (1.08 g, 1.87 mmol). The reaction was heated to 100° C. for 16hr under N₂ atmosphere. After cooling the reaction to room temperature,water was added and extracted with EA (50 mL×3). The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄. Afterfiltration, the filtrate was concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography, eluted withPE/EA to afford the desired product (2 g, 35%) as a yellow solid. ESI-MSm/z: 303.00 [M+H]⁺.

A solution of the compound from step a (1.5 g) and TFA (8 mL) in DCM (20mL) was stirred for 2 hr at room temperature. The residue product waspurified by reverse phase flash with the following conditions (water(0.05% FA)/MeOH) to afford the desired product (520 mg) as a whitesolid. ESI-MS m/z: 203.00 [M+H]⁺.

Intermediate 87 Steps c-e

The following example was prepared in an analogous fashion toIntermediate 86 to afford the desired product (300 mg, 86%) as a yellowsolid. ESI-MS m/z: 192.95 [M+H]⁺.

Intermediate 88

Intermediate 88 Step a

A solution of 6-bromo-3-methoxypyridin-2-amine (1 g, 4.93 mmol) and2-bromo-1-cyclopropylethanone (963 mg, 5.91 mmol) in EtOH (10 mL) wasstirred for overnight at 80° C. The reaction was quenched with water andthe resulting mixture was extracted with EA (3×200 mL). The combinedorganic layers were washed with NaHCO₃ (3×200 mL), dried over anhydrousNaSO₄. After filtration, the filtrate was concentrated under reducedpressure. The residue was purified by silica gel column chromatography,eluted with PE/EA to afford the desired product (360 mg, 27%) as ayellow solid. ESI-MS m/z: 267.00 [M+H]⁺.

Intermediate 88 Steps b and c

The following example was prepared in an analogous fashion toIntermediate 86 to afford the desired product (110 mg, 62%) as a yellowsolid. ESI-MS m/z: 233.00 [M+H]⁺.

Intermediate 89

The following example was prepared in an analogous fashion toIntermediate 86 to afford the desired product. ESI-MS m/z: 206.95[M+H]⁺.

Intermediate 90

Intermediate 90 Step a

A solution/mixture of 3-cyclopropoxy-2-nitropyridine (1.8 g, 1 mmol), Fe(5.58 g, 100 mmol) and NH₄C₁ (5.34 g, 100 mmol) in EtOH (60 mL) and H₂O(30 mL) was stirred for 2 hr at 80° C. The residue was purified bysilica gel column chromatography, eluted with PE/EA to afford thedesired product (1.4 g, 93%) as a yellow solid. ESI-MS m/z: 151.00[M+H]⁺.

Intermediate 90 Step b

A solution/mixture of the compound from step a (1.4 g, 9.32 mmol) andBr₂ (2.98 g, 18.64 mmol) in AcOH (30 mL) was stirred for overnight atroom temperature. The resulting mixture was quenched with sat. sodiumthiosulfate and extracted with EA. The combined organic layers werewashed with NaHCO₃ solution (3×200 mL), dried over anhydrous Na₂SO₄.After filtration, the filtrate was concentrated under reduced pressureto afford the desired product (1.4 g, 66%) as a yellow solid. ESI-MSm/z: 229.00 [M+H]⁺.

Intermediate 90 Steps c-e

The following example was prepared in an analogous fashion toIntermediate 86 to afford the desired product. ESI-MS m/z: 219.00[M+H]⁺.

Intermediate 91

Intermediate 91 Steps a and b

A solution of 3-hydroxy-4-aminobenzoic acid (10 g, 65.30 mmol) andmethacrolein (9.15 g, 130.60 mmol) in HCl (40 mL) and AcOH (60 mL) wasstirred for 1 hr at 100° C. under N₂ atmosphere. The combined layerswere concentrated under reduced pressure to afford the desired product(5.6 g, 42%) as a brown solid. ESI-MS m/z: 204.00 [M+H]⁺.

A solution/mixture of the compound from step a (5.6 g, 27.56 mmol) andH₂SO₄ (5 mL) in MeOH (50 mL,) was stirred for 1 hr at 80° C. underreflux. The mixture was neutralized to pH 7 with NaOH solution. Theresulting mixture was extracted with EA (3×300 ml). The combined organiclayers were concentrated under reduced pressure to afford the desiredproduct (2.5 g, 42%) as yellow solid. ESI-MS m/z: 218.00 [M+H]⁺.

Intermediate 91 Steps c and d

A solution of the compound from step b (500 mg, 2.30 mmol),2-bromo-1,1-difluoroethane (667 mg, 4.6 mmol) and K₂CO₃ (954 mg, 6.91mmol) in DMF (20 mL) was stirred for overnight at 80° C. The residue waspurified by silica gel column chromatography, eluted with PE/EA toafford the desired product (500 mg, 62%) as a yellow solid. ESI-MS m/z:282.00 [M+H]⁺.

A solution of the compound from step c (500 mg, 1.78 mmol) and LiOH (426mg, 17.78 mmol) in MeOH (20 mL) and H₂O (20 mL) was stirred for 30 minat room temperature. The mixture was acidified to pH 7 with HCl. Thecrude product was re-crystallized from the water to afford the desiredproduct (270 mg, 57%) as a yellow solid. ESI-MS m/z: 267.95 [M+H]⁺.

Intermediate 92 Steps a and b

A solution of methyl 8-hydroxy-3-methylquinoline-6-carboxylate (3 g,13.81 mmol), sodium 2-chloro-2,2-difluoroacetate (3.16 g, 20.72 mmol)and Cs₂CO₃ (9 g, 27.61 mmol) in DMF (30 mL) was stirred for 4 hr at 80°C. under N₂ atmosphere. The resulting mixture was extracted with EA(3×200 mL). The combined organic layers were washed with water (3×200ml), dried over anhydrous Na₂SO₄. After filtration, the filtrate wasconcentrated under reduced pressure. The residue was purified by silicagel column chromatography, eluted with PE/EA to afford the desiredproduct (2 g, 54%) as a yellow solid. ESI-MS m/z: 268.00 [M+H]⁺.

A solution of the compound from step a (3 g, 11.23 mmol), LiOH (2.7 g,112.26 mmol) in MeOH (30 mL) and H₂O (30 mL) was stirred for 4 hours atroom temperature. The mixture/residue was acidified to pH 7 with HCl,the resulting mixture was filtered, the filter cake was washed withwater. The residue was purified by reverse flash chromatography with thefollowing conditions (silica gel, MeCN in water, 10% to 50% gradient in10 min) to afford the desired product (800 mg, 28%) as a yellow solid.ESI-MS m/z: 254.20 [M+H]⁺.

Intermediate 93 Steps a and b

A mixture of methyl 4-amino-3-chlorobenzoate (1.5 g, 8.08 mmol) and(2Z)-2-chlorobut-2-enal (1.27 g, 12.12 mmol) in HCl (10 mL), AcOH (15mL) was stirred for 20 min at 100° C. The reaction was monitored byLCMS. The mixture was allowed to cool down to room temperature,concentrated under vacuum. The residue dissolved in MeOH (20 mL) andH₂SO₄ (2 mL), stirred for another 1 h at 80° C. The resulting mixturewas poured into water, extracted with EA. The combined organic layerswere concentrated under reduced pressure. The residue was purified byreverse flash chromatography to afford the desired product (110 mg, 5%)as a yellow solid. ESI-MS m/z: 269.90 [M+H]⁺.

Into a 100 mL round-bottom flask were added the compound from step a(110 mg, 0.40 mmol), THE (4 mL), MeOH (1 mL), LiOH (97 mg, 4.07 mmol)and H₂O (1 mL) at room temperature. The resulting mixture was stirredfor 3 hours at room temperature. The mixture was acidified to pH 5 withHCl (1 M aq.). The product was precipitated. The precipitated solidswere collected by filtration and washed with water to afford the desiredproduct (65 mg). ESI-MS m/z: 256.00 [M+H]⁺.

Intermediate 94

Intermediate 94 Step a

A solution of methyl 3-bromo-5-methoxy-4-nitrobenzoate (3.60 g, 12.43mmol) in toluene (20 mL) was treated withtributyl(1-ethoxyethenyl)stannane (5.39 g, 14.91 mmol) and Pd(dppf)Cl₂(1.82 g, 2.49 mmol) and stirred for 2 hours at 110° C. under nitrogenatmosphere. The reaction was monitored by LCMS. The resulting mixturewas filtered, the filter cake was washed with EA. The filtrate wasconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (eluted with 25% ethyl acetate in hexanes) toafford the compound (2.69 g, 77%) as a yellow oil. ESI-MS m/z: 282.20[M+H]⁺.

Intermediate 94 Step b

A solution of the compound from step a (2.69 g, 9.56 mmol) in DCM (30mL) was treated with HCl (3 mL) for overnight at room temperature. Thereaction was monitored by LCMS. The aqueous layer was extracted withCH₂Cl₂. The residue was purified by silica gel column chromatography(eluted with 50% ethyl acetate in hexanes) to afford the compound (1.82g, 75%) as a white solid. ESI-MS m/z: 253.85 [M+H]⁺.

Intermediate 94 Steps c and d

Into a 250 mL round-bottom flask were added the compound from step b(1.82 g, 7.18 mmol) and a solution of NaOAc (21.83 g, 266.09 mmol) inMeOH (90 mL) and THE (90 mL) was treated with SnCl₂ (18.19 g, 94.92mmol) and stirred for 16 hrs at room temperature under nitrogenatmosphere. The reaction was monitored by LCMS. The resulting mixturewas filtered, the filter cake was washed with ethyl acetate. Thefiltrate was concentrated under reduced pressure. The residue waspurified by silica gel column chromatography, eluted with PE/EA (7:3) toafford the compound (851 mg, 54%) as a white solid. ESI-MS m/z: 222.10[M+H]⁺. Into a 250 mL round-bottom flask were added the compound fromstep c (840 mg, 3.80 mmol) and THE (40 mL) at room temperature. Asolution of LiOH (909 mg, 37.97 mmol) in MeOH (10 mL) and H₂O (10 mL)was added and stirred for 5 hours at room temperature. The reaction wasmonitored by LCMS. The mixture was acidified to pH 4 with HCl then theproduct was precipitated. The precipitated solids were collected byfiltration and washed with H₂O to afford the compound (646 mg, 81%) asan orange solid. ESI-MS m/z: 207.95 [M+H]⁺.

Intermediate 95

The following compounds was prepared in an analogous procedure toIntermediate 94 to afford the desired product (741 mg, 88% o) as anorange solid. ESI-MS m/z: 178.10 [M+H]⁺.

The following Table 5 contains examples that were prepared with thesimilar method to Example 1 Step c (PyBOP or HATU). The majority ofcompounds were purified by prep-HPLC (ACN/H₂O, 20-90%, 25 min), and somewere purified by automated column chromatography (silica gel). The arylacid coupling partners were prepared according to Intermediates 1-95, orby analogous procedures with slight modifications and also preparedaccording to procedures found in U.S. patent application Ser. No.16/930,622.

TABLE 5 MS⁺ Example Structure m/z 491

646.27 492

590.19 493

635.34 494

669.12 495

607.22 496

641.17 497

683.19 498

621.25 499

655.15 500

642.95 501

653.29 502

625.39 503

659.13 504

702.24 505

667.32 506

669.43 507

650.42 508

676.58 509

667.22 510

648.29 511

667.22 512

689.42 513

639.28 514

695.48 515

689.39 516

687.22 517

637.18 518

693.20 519

687.42 520

699.18 521

697.12 522

668.22 523

671.41 524

669.33 525

621.51 526

641.49 527

641.41 528

632.23 529

658.20 530

667.49 531

611.43 532

671.30 533

638.47 534

604.51 535

620.47 536

620.48 537

640.48 538

669.46 539

637.45 540

693.51 541

683.49 542

687.45 543

697.32 544

664.42 545

636.48 546

680.40 547

652.57 548

664.40 549

636.45 550

684.40 551

676.38 552

685.35 553

687.36 554

638.49 555

656.48 556

648.17 557

657.43 558

659.42 559

642.39 560

664.49 561

687.27 562

656.21 563

648.17 564

642.16 565

657.19 566

659.20 567

702.31 568

674.46 569

712.42 570

652.20 571

636.26 572

677.32 573

627.60 574

630.02 575

620.08 576

664.97 577

665.03 578

674.46 579

698.02 580

707.96 581

717.45 582

665.96 583

656.02 584

675.45 585

630.02 586

639.96 587

665.45 588

684.88 589

675.39 590

599.54 591

685.51 592

645.54 593

641.72 594

651.66 595

661.15 596

689.08 597

695.69 598

669.65 599

687.64 600

679.59 601

659.05 602

719.51 603

715.12 604

659.71 605

713.68 606

705.63 607

685.75 608

629.68 609

674.07 610

727.71 611

737.65 612

719.66 613

683.99 614

675.94 615

710.03 616

724.06 617

734.00 618

716.01 619

670.42 620

685.43 621

715.86 622

646.13 623

620.16 624

637.54 625

631.58 626

653.99 627

666.03 628

619.55 629

677.70 630

669.65 631

664.12 632

659.71 633

638.90 634

620.83 635

627.96 636

628.10 637

631.25 638

665.01 639

676.38 640

695.05 641

695.05 642

574.48 643

593.48 644

615.53 645

563.56 646

560.52 647

582.55 648

584.51 649

632.46 650

666.52 651

672.51 652

638.45 653

600.46 654

666.42 655

664.44 656

630.40 657

630.40 658

648.43 659

656.47 660

687.28 661

638.47 662

648.45 663

656.46 664

689.24 665

659.42 666

657.38 667

630.40 668

642.38 669

682.35 670

692.32 671

700.33 672

731.15 673

686.24 674

701.30 675

671.16 676

654.35 677

664.19 678

672.31 679

703.06 680

658.24 681

673.25 682

643.26 683

638.33 684

648.27 685

656.33 686

642.29 687

657.27 688

627.26 689

687.13 690

608.32 691

574.17 692

558.13 693

573.01 694

588.16 695

614.17 696

572.99 697

676.45 698

656.31 699

574.15 700

636.28 701

606.28 702

600.47 703

620.08 704

599.95 705

604.46 706

604.47 707

623.47 708

600.53 709

584.40 710

625.41 711

614.51 712

666.53 713

687.48 714

638.50 715

659.48 716

666.57 717

685.46 718

638.53 719

657.39 720

676.50 721

648.50 722

611.46 723

629.48 724

595.44 725

684.45 726

618.52 727

657.40 728

577.43 729

640.49 730

661.43 731

659.47 732

638.48 733

659.59 734

657.41 735

638.46 736

659.45 737

657.36 738

593.50 739

649.42 740

575.49 741

603.47 742

621.47 743

658.45 744

650.40 745

656.44 746

677.40 747

675.37 748

666.40 749

672.43 750

693.39 751

682.39 752

656.39 753

677.38 754

666.37 755

608.38 756

622.40 757

644.36 758

656.36 759

660.29 760

675.25 761

648.34 762

572.38 763

598.38 764

662.31 765

688.32 766

666.28 767

707.08 768

707.08 769

677.12 770

739.09 771

649.18 772

667.21 773

696.53 774

685.27 775

684.46 776

676.49 777

711.39 778

717.05 779

655.27 780

670.30 781

687.41 782

666.33 783

676.35 784

685.24 785

717.22 786

657.24 787

638.38 788

648.34 789

689.14 790

642.37 791

656.39 792

627.36 793

636.50 794

654.37 795

640.46 796

646.56 797

655.41 798

625.41 799

684.24 800

688.11 801

694.11 802

702.27 803

733.11 804

703.18 805

558.16 806

558.16 807

558.16 808

578.11 809

562.14 810

612.14 811

596.18 812

602.19 813

600.17 814

628.13 815

590.18 816

648.21 817

652.16 818

696.19 819

646.1 820

632.18 821

630.19 822

674.23 823

658.18 824

618.20 825

667.16 826

648.21 827

628.14 828

648.21 829

668.23 830

624.20 831

618.20 832

602.17 833

620.16 834

636.13 835

666.21 836

638.19 837

694.16 838

682.18 839

638.19 840

638.19 841

654.16 842

654.16 843

654.16 844

654.16 845

688.18 846

692.22 847

684.20 848

654.19 849

670.16 850

632.18 851

632.18 852

642.15 853

642.15 854

651.13 855

651.13 856

662.13 857

666.21 858

670.16 859

684.20 860

676.17 861

685.16 862

656.18 863

666.21 864

670.16 865

684.20 866

676.17 867

685.16 868

656.18 869

715.09 870

715.09 871

654.20 872

620.40 873

620.15 874

614.20 875

614.20 876

630.20 877

672.20 878

664.10 879

614.25 880

648.15 881

589.15 882

589.15 883

637.15 884

671.00 885

659.10 886

693.05 887

559.10 888

589.10 889

682.10

Intermediate 96

Intermediate 96 Step a

Into a 100 mL round-bottom flask were added 5-bromoisatin (1 g, 4 mmol),and DCM (5 mL) at room temperature. DAST (1.43 g, 9 mmol) was addeddropwise. The resulting mixture was stirred for 1 hr at 0° C. undernitrogen atmosphere. The reaction was monitored by LCMS. The reactionwas quenched with water and the aqueous layer was extracted with EA. Theresidue was purified by silica gel column chromatography (eluted with25% ethyl acetate in hexanes) to afford the desired compound (900 mg,82%) as a light-yellow solid.

Intermediate 96 Step b

A solution of the compound from step a (300 mg, 1.2 mmol),bis(pinacolato)diboron (461 mg, 1.8 mmol), Pd(dppf)Cl₂CH₂Cl₂ (197 mg,0.24 mmol), KOAc (356 mg, 4 mmol) in dioxane (5 mL) mixture was stirredfor 2 hr at 90° C. under nitrogen atmosphere. The reaction was monitoredby TLC. The aqueous layer was extracted with EtOAc. The resultingmixture was concentrated under reduced pressure. The residue waspurified by reverse flash to afford the desired compound (297 mg, 83%)as yellow solid.

Intermediate 97

The following compound was prepared in an analogous fashion toIntermediate 96 Step b above to afford the desired product that was useddirectly in the next step crude.

Intermediate 98

In a 50 mL round bottom flask, to a solution of1-bromo-2-chloro-3,4-difluorobenzene (850 mg, 3.7 mmol) in THE (30 mL)was added n-BuLi (2.8 mL, 5.6 mmol) dropwise at −78° C. under N₂atmosphere. The reaction mixture was stirred at −78° C. for 5 mins. ThenB(OMe)₃ (583 mg, 5.6 mmol) was added dropwise and the mixture andstirred for another 30 mins at room temperature. The mixture wasacidified to pH 5 with HCl (3 M aq.). The reaction was quenched withsat. NH₄Cl, and then the mixture was extracted with EtOAc. The combinedorganic extracts were washed with brine, dried over anhydrous Na₂SO₄,and concentrated under vacuum to afford the desired crude compound (790mg, 109%) as a yellow oil. The crude product was used in the next stepdirectly without further purification.

Example 890

Example 890 Step a

To a 250 mL round-bottom flask containing Intermediate 36 Step c (5.09g, 14.50 mmol) was added THF (24.16 mL), Acetone (24.16 mL) and Water(24.16 mL). The flask was cooled to 0° C., and NMO (4.25 g, 36.2 mmol)was added followed by potassium osmate dihydrate (0.230 g, 0.623 mmol).The reaction was stirred for 10 minutes, warmed to room temperature andallowed to stir overnight for 20 hr. Sodium sulfite was added, themixture diluted with water and the reaction stirred for 20 minutes. Themixture was diluted with EtOAc and the aqueous layer was extracted. Thecombined organics were dried with sodium sulfate, filtered, andconcentrated. The crude mixture was purified by automated columnchromatography (silica gel, 0-100% ethyl acetate in hexanes) to affordthe desired product as a white solid (4.12 g, 74%) and mixture ofdiastereomers. ESI-MS m/z: 385.21 [M+H]⁺.

Example 890 Step b

To a 250 mL round-bottom flask containing step a (4.118 g, 10.69 mmol)was added DCM (36 mL, 0.3 M). DMAP (0.065 g, 0.535 mmol) was addedfollowed by TEA (4.47 ml, 32.1 mmol). The flask was cooled to 0° C., andTsCl (2.242 g, 11.76 mmol) was added, stirred for 10 minutes, warmed toroom temperature and monitored by LCMS (1 hr). The reaction wasconcentrated, and the crude residue was purified by automated columnchromatography (silica gel, 0-100% ethyl acetate in hexanes) to affordthe desired product as a white solid (3.92 g, 99%), a mixture ofdiastereomers, and a mixture of epoxide (major) and tosylate. ESI-MSm/z: 367.19 [M+H]⁺.

Example 890 Step c

To a 500 mL round-bottom flask containing step b (3.92 g, 10.70 mmol)was added ammonia (239 mL, 1670 mmol, 7N in MeOH) at 0° C. The reactionwas stirred for 10 minutes, warmed to room temperature and monitored byLCMS (3.5 hr). The stir was then removed and the reaction concentrated.The crude material was dissolved in EtOAc and the organics were washed3× with saturated sodium bicarbonate. The organics were concentrated,and the crude material was triturated with DCM/hexanes to afford a whitesolid (2.41 g, 59%) as a mixture of diastereomers. ESI-MS m/z: 384.21[M+H]⁺.

Example 890 Step d

To a 100 mL round bottom flask equipped with a stir bar was added step c(1.000 g, 2.60 mmol) and2-cyclopropyl-7-methoxy-2H-indazole-5-carboxylic acid (0.605 g, 2.60mmol). The solids were dissolved in N,N-Dimethylformamide (13.02 mL, 0.2M), and DIPEA (0.909 mL, 5.21 mmol) was added. The vial was cooled to 0°C., and PyBOP (1.626 g, 3.12 mmol) was added. The reaction was stirredfor 10 minutes, warmed to room temperature and monitored by LCMS (1 hr).The reaction was quenched with sat. ammonium chloride and diluted withEtOAc. The aqueous was extracted with EtOAc, and the combined organicswere dried with sodium sulfate, filtered, and concentrated. The crudemixture was purified by automated column chromatography (silica gel,0-100% ethyl acetate in hexanes) to afford the desired product as anoil. The oil was dissolved in EtOAc and washed with water and brine toremove DMF and concentrated to afford the product as a white solid (597mg, 38%) as a mixture of diastereomers. ESI-MS m/z: 598.12 [M+H]⁺.

Example 891

To a 2 dram vial equipped with a stir bar and septa cap was addedPdCl₂(dppf) (6.11 mg, 8.36 μmol), potassium carbonate (26.0 mg, 0.188mmol), (4-cyclopropylphenyl)boronic acid (16.24 mg, 0.100 mmol) andExample 890 (50 mg, 0.084 mmol). The material was dissolved in1,4-dioxane (0.3 mL, 4:1, 0.2 M) and water (0.084 mL) and purged withnitrogen. The reaction was heated to 90° C. and monitored by LCMS (2hr). The reaction was diluted with EtOAc, filtered through a short padof silica gel, the pad was washed with EtOAc and the organics wereconcentrated. The crude material was purified and diastereomers wereseparated by prep-HPLC (ACN/H₂O, 20-90%, 25 min) to afford the desiredproduct as a white solid (3 mg, 6%). ESI-MS m/z: 636.26 [M+H]⁺.

Example 892

The following example was purified by prep-HPLC (ACN/H₂O, 20-90%, 25min) from Example 891 as a white solid (3 mg, 6%). ESI-MS m/z: 636.26[M+H]⁺.

The following Table 6 contains examples that were prepared with asimilar method to Example 891 using Example 890. If necessary to pushconversion, more palladium and boronic acid were added. The majority ofcompounds were purified and the diastereomers were separated byprep-HPLC (ACN/H₂O, 20-90%, 25 min). If reported as a mixture ofdiastereomers, they were likely not separable by HPLC.

TABLE 6 MS⁺ Example Structure m/z 893

676.19 894

676.19 895

646.22 896

646.22 897

680.08 898

680.08 899

666.01 900

632.08 901

632.08 902

651.06 903

600.06 904

668.07 905

656.16 906

656.16 907

640.22 908

640.22 909

680.21 910

614.15 911

614.15 912

614.24 913

644.40 914

644.40 915

650.21 916

650.16 917

662.43 918

662.43 919

680.45 920

680.45 921

648.19 922

648.19 923

626.44 924

626.44 925

662.44 926

662.44 927

698.12 928

676.44 929

666.41 930

666.47 931

666.42 932

666.09 933

660.05 934

660.05 935

664.46 936

669.65 937

686.10 938

685.61 939

684.62 940

684.62 941

685.61 942

715.64 943

651.66 944

669.65 945

651.61 946

669.60 947

651.61 948

651.61 949

651.61 950

670.59 951

670.59 952

666.00 953

726.11 954

726.11 955

712.18 956

712.18 957

712.08 958

712.08 959

694.06 960

694.06 961

666.14 962

662.19 963

666.14 964

650.17 965

646.20 966

647.25 967

661.20 968

602.15 969

635.25 970

635.30 971

651.25 972

698.15 973

654.20 974

682.20 975

630.20 976

652.25 977

636.25 978

680.20 979

676.25 980

622.25 981

661.25 982

620.15 983

714.15 984

636.25 985

632.20 986

648.10 987

648.15 988

632.15 989

648.15 990

648.10 991

648.10 992

632.15 993

648.10 994

670.20 995

687.25 996

668.15 997

650.15

The following Table 7 contains examples that were prepared with thesimilar method to Example 1 Step c (PyBOP or HATU). The majority ofcompounds were purified by prep-HPLC (ACN/H₂O, 20-90%, 25 min) Ifdepicted as a single diastereomer, they were separated during prep-HPLCpurification. The amine coupling partners were prepared in analogousfashion to Intermediates 45 & 46. The aryl acid coupling partners wereprepared according to Intermediates 1-95, or by analogous procedureswith slight modifications and also prepared according to proceduresfound in U.S. patent application Ser. No. 16/930,622.

TABLE 7 Example Structure MS⁺ m/z 998

650.16 999

650.16 1000

669.12 1001

669.12 1002

684.15 1003

684.15 1004

703.11 1005

703.11 1006

671.27 1007

671.27 1008

705.25 1009

705.25 1010

684.40 1011

684.40 1012

684.45 1013

684.45 1014

703.44 1015

703.44 1016

701.21 1017

701.21 1018

675.28 1019

675.28 1020

705.42 1021

705.42 1022

622.49 1023

622.49 1024

656.43 1025

656.43 1026

643.43 1027

643.43 1028

677.42 1029

677.42 1030

641.42 1031

641.42 1032

675.13 1033

675.13 1034

712.48 1035

712.48 1036

731.47 1037

731.47 1038

686.44 1039

686.44 1040

705.41 1041

705.41 1042

702.26 1043

733.36 1044

694.22 1045

666.19 1046

684.95 1047

684.90 1048

641.99 1049

641.99 1050

666.05 1051

666.06 1052

666.08 1053

601.52 1054

601.52 1055

623.40 1056

623.51 1057

625.50 1058

625.48 1059

604.54 1060

604.57 1061

614.47 1062

614.88 1063

604.59 1064

604.53 1065

640.12 1066

640.02

Intermediate 99

Intermediate 99 Steps a and b

5-(benzyloxy)-2-(hydroxymethyl)-1H-pyridin-4-one (5 g, 21.62 mmol),Trt-Cl (6.63 g, 23.78 mmol) and DMAP (2.91 g, 23.78 mmol) was suspendedin DMF (50 mL), and the mixture was stirred for overnight at 95° C. Thereaction was monitored by LCMS. After cooling to room temperature, themixture was treated with cold water and stirred for 30 min. Theprecipitated solids were collected by filtration and washed with waterand MeOH, dried to give the desired product (8.8 g, 86%) as a light greysolid. ESI-MS m/z: 474.30 [M+H]⁺.

To a solution of the compound from step a (4.4 g, 9.29 mmol) in THE (120mL), H₂O (12 mL) and aqueous NaOH 2 M (6 mL) was added Pd/C (2.2 g,20.67 mmol) under nitrogen atmosphere. The resulting mixture was stirredfor 2 hours at room temperature under hydrogen atmosphere. The reactionwas monitored by TLC. The resulting mixture was filtered, the filtercake was washed with acetonitrile. The filtrate was concentrated underreduced pressure. To a suspension of the residue in DCM (100 mL) wereadded DMAP (2.27 g, 18.58 mmol) and thiophosgene (1.58 g, 13.75 mmol).After stirring for 1 hour at room temperature, the reaction wasmonitored by TLC. The reaction was quenched with water and extractedwith CH₂Cl₂. The combined organic layers were washed with brine, driedover anhydrous Na₂SO₄. After filtration, the filtrate was concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography, eluted with P/EA (10:1) to afford the desired product(3.3 g, 83%) as a yellow solid. ESI-MS m/z: 448.20 [M+Na]⁺.

Intermediate 99 Steps c and d

To a solution of the compound from step b (3.3 g, 7.75 mmol) in DCM (100mL) were added HF Pyridine (33 mL) and DBDMH (3.77 g, 13.18 mmol)keeping the temperature below −60° C. The reaction mixture was allowedto warm to 0° C. over 20 min. After stirring for 2 hours at thetemperature, the reaction was quenched with 2 M aqueous NaOH, extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure. The residue was purified by silica gel column chromatography,eluted with PE/EA (1:1) to afford the desired product (740 mg, 50%) as ayellow oil. ESI-MS m/z: 190.05 [M+H]⁺.

Intermediate 99 Step e

To a solution of the compound from step d (550 mg, 2.94 mmol) in acetone(20 mL) and H₂O (10 mL) were added 2-methyl-2-butene (2.06 g, 29.40mmol), NaH₂PO₄ (529 mg, 4.41 mmol) and NaClO₂ (532 mg, 5.88 mmol). Theresulting mixture was stirred for 2 hours at room temperature. Themixture was treated with NaHSO₃ (1.07 g, 10.28 mmol) and concentratedunder reduce pressure to remove acetone. The mixture was treated withbrine (10 mL) and extracted twice with EA/THF (1:1). The combinedorganic layers were dried over anhydrous Na₂SO₄. After filtration, thefiltrate was concentrated under reduced pressure. The residue waspurified by reverse flash chromatography to afford the desired product(151.8 mg, 25%) as a white solid. ESI-MS m/z: 203.90 [M+H]⁺.

Intermediate 100

Intermediate 100 Steps and b

A mixture of 5-bromo-7-methoxy-1H-indazole (500 mg, 2.2 mmol), Cs₂CO₃(2.15 g, 6.6 mmol) and CBr₂F₂ (5 mL) in ACN (25 mL) was stirred forovernight at room temperature under N₂ atmosphere. The residue waspurified by reverse flash chromatography to afford the desired product(200 mg, 26%) as a yellow oil. ESI-MS m/z: 355.00 [M+H]⁺.

A solution of the compound from step a (400 mg, 1.12 mmol) and AgBF₄(656 mg, 3.37 mmol) in DCM (20 mL) was stirred for 2 hours at roomtemperature. The resulting mixture was washed with 3×100 mL of NaHCO₃.The resulting mixture was extracted with EA (3×100 mL). The combinedorganic layers were washed with water (3×100 mL), dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure. The residue was purified by silica gel column chromatography,eluted with PE/EA to afford the desired product (300 mg, 90%) as ayellow solid. ESI-MS m/z: 295.00 [M+H]⁺.

Intermediate 100 Steps c and d

The following compound was prepared according to the same procedures asIntermediate 77 steps d and e to afford the desired product (200 mg,72%) as a white solid. ESI-MS m/z: 275.00 [M+H]⁺.

Intermediate 101

Intermediate 101 Steps a, b and c

A mixture of 2-amino-5-bromo-3-methoxybenzaldehyde (9.6 g, 41.73 mmol)and urea (37.59 g, 625.92 mmol) was stirred for 2 hours at 180° C. Themixture was allowed to cool down to room temperature and poured into icewater. The precipitated solids were collected by filtration and washedwith water and dried in vacuum to afford the crude product (11 g) as agrey solid. ESI-MS m/z: 254.85 [M+H]⁺.

A solution of the compound from step a (11 g, 43.12 mmol) in phosphorusoxychloride (100 mL) was stirred for 5 hours at 110° C. The mixture wasallowed to cool down to room temperature and concentrated under reducedpressure. The residue was dissolved in EA and poured into ice water withvigorous stirring. The resulting mixture was extracted with EA. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄. After filtration, the filtrate was concentrated under reducedpressure. The residue was purified by silica gel column chromatography,eluted with PE/EA (2:1) to afford the desired product (2.4 g, 20%) as alight-yellow solid. ESI-MS m/z: 272.95 [M+H]⁺.

A mixture of the compound from step b (2.4 g, 8.77 mmol) and NaOMe (0.47g, 8.77 mmol) in MeOH (30 mL) was stirred for 1 h at room temperatureand heated to reflux for 1 hour. The reaction was monitored by LCMS. Theresulting mixture was concentrated under reduced pressure. The residuewas stirred with water. The precipitated solids were collected byfiltration and washed with water and dried in vacuum to afford thedesired product (2.2 g, 93%) as a light-yellow solid. ESI-MS m/z: 268.85[M+H]⁺.

Intermediate 101 Steps d and e

The following compound was prepared according to the same procedures asIntermediate 77 steps d and e to afford the desired product (658.6 mg,77%) as a white solid. ESI-MS m/z: 235.00 [M+H]⁺.

Intermediate 102

Intermediate 102 Steps a and b

A solution of methyl 4-amino-3-methoxybenzoate (4 g, 22.07 mmol) in HCl(15 mL) were added methacrolein (3.87 g, 55.19 mmol). The resultingmixture was stirred for 5h at 100° C. The reaction was monitored byLCMS. The resulting mixture was concentrated under reduced pressure. Thecrude product was used in the next step directly without furtherpurification. This resulted in 8-methoxy-3-methylquinoline-6-carboxylicacid (4 g, 83%) as a brown crude solid. ESI-MS m/z: 218.05 [M+H]⁺.

A solution of the compound from step a (4 g, 18.41 mmol) in MeOH (120mL) were added SOCl₂ (8.76 g, 73.65 mmol). The resulting mixture wasstirred for 40 min at 80° C. The reaction was monitored by LCMS. Theresulting mixture was concentrated under reduced pressure. The residuewas dissolved in water (40 mL). The mixture was basified to pH 7 with 1Maq NaHCO₃, extracted and evaporated. The residue was purified by silicagel column chromatography, eluted with PE/EA (1:2) to afford thecompound (1.8 g, 42%) as a brown solid. ESI-MS m/z: 232.00 [M+H]⁺.

Intermediate 102 Steps c and d

A solution of the compound from step b (1.8 g, 7.78 mmol) in DCM (20 mL)was added with m-CPBA (4.03 g, 23.35 mmol) in portions at 0° C. Theresulting mixture was stirred for overnight at room temperature. Thereaction was monitored by LCMS. The resulting mixture was washed with 1M NaOH. The aqueous layer was basified to pH 2 with conc. HCl. Theresulting mixture was filtered, the filter cake was washed with water.The filtrate was concentrated under reduced pressure. The crude productwas used in the next step directly without further purification. ESI-MSm/z: 233.95 [M+H]⁺.

A solution of the compound from step c (5 g crude) and POCl₃ (42.73 g,278.707 mmol, 13 equiv) was stirred for 1 h at 95° C. The reactionmixture was slowly added to water (20 mL) and the reaction was monitoredby LCMS. The resulting mixture was concentrated under reduced pressure.The residue was purified by reverse flash to afford the titled compound(100 mg, 1.85%) as a white solid. ESI-MS m/z: 251.95 [M+H]⁺.

Intermediate 103

This intermediate was prepared in an analogous fashion to Intermediate102. (80 mg, 70%) ESI-MS m/z: 277.95 [M+H]⁺.

Intermediate 104

The following compound was prepared in an analogous fashion toIntermediate 102 above (64 mg, 74%) ESI-MS m/z: 318.10 [M+H]⁺.

Intermediate 105

The above compound was prepared in an analogous fashion to Intermediate102 above (32 mg, 46%) ESI-MS m/z: 268.15 [M+H]⁺.

Intermediate 106

Intermediate 106 Steps a, b and c

Into a 250 mL round-bottom flask were added2,5-dichloro-3-fluoropyridine (10 g, 60.25 mmol),(4-methoxyphenyl)methanol (9.16 g, 66.27 mmol), Cs₂CO₃ (39.26 g, 121mmol) and DMF (40 mL) at room temperature. The resulting mixture wasstirred for 1h at 80° C. The reaction was monitored by LCMS. The aqueouslayer was extracted with EtOAc. The resulting mixture was washed withbrine. The resulting mixture was concentrated under vacuum. The residuewas purified by silica gel column chromatography to afford the desiredcompound (14 g, 81%) as a white solid. ESI-MS m/z: 284.25 [M+H]⁺.

Into a 100 mL round-bottom flask were added the compound from the step a(5 g, 18 mmol) and DCM (24 mL) at room temperature. To the above mixturewas added CF₃COOH (6 g, 52.62 mmol) dropwise. The resulting mixture wasstirred for additional overnight at room temperature. The reaction wasmonitored by LCMS. The aqueous layer was extracted with CH₂Cl₂. Theresulting mixture was concentrated under vacuum and the crude productwas used in the next step directly without further purification. ESI-MSm/z: 165.10 [M+H]⁺.

Into a 100 mL round-bottom flask were added the compound from the step c(6.2 g, 37.81 mmol), 12 (10.56 g, 41.59 mmol), K₂CO₃ (10.45 g, 75.61mmol) and H₂O (40 mL) at room temperature. The resulting mixture wasstirred for 2h at room temperature. The reaction was monitored by LCMS.The reaction was quenched with sat. sodium hyposulfite (aq.) at 0° C.The mixture was acidified to pH 6 with conc. HCl. The aqueous layer wasextracted with EtOAc. The residue was purified by silica gel columnchromatography to afford the desired compound as a white solid. ESI-MSm/z: 290.15 [M+H]⁺.

Intermediate 106 d and e

Into a 250 mL round-bottom flask were added the compound from the step c(3.9 g, 13.45 mmol), (2-methyloxiran-2-yl)methyl4-methylbenzenesulfonate (3.91 g, 16.14 mmol), KI (2.2 g, 13.45 mmol),K₂CO₃ (3.7 g, 26.91 mmol) and DMF (20 mL) at room temperature. Theresulting mixture was stirred overnight at 50° C. The reaction wasmonitored by LCMS. The reaction was quenched with sat. NH₄Cl (aq.) at 0°C. The aqueous layer was extracted with EtOAc. The residue was purifiedby reverse flash chromatography to afford the desired compound (3.0 g,65%) as a yellow oil. ESI-MS m/z: 359.95 [M+H]⁺.

Into a 50 mL 3-necked round-bottom flask were added the compound fromthe step d (1.85 g, 5.13 mmol) and LDA (2M in THF) (3.9 mL) at 0° C. Theresulting mixture was stirred for 1h at room temperature under nitrogenatmosphere. The reaction was monitored by LCMS. Desired product could bedetected by LCMS. The reaction was quenched with sat. NH₄Cl (aq.) andextracted with EA, The resulting mixture was concentrated under reducedpressure. The residue was purified by reverse flash chromatography toafford the desired compound as a yellow oil. ESI-MS m/z: 360.15 [M+H]⁺.

Intermediate 106 Steps f and g

Into a 250 mL round-bottom flask were added the compound from the step e(6.6 g, 18.33 mmol), acetone (20 mL) and Jones reagent (8 mL, 40.39mmol) in portions at 0° C. under nitrogen atmosphere. The resultingmixture was stirred for 3 h at room temperature under nitrogenatmosphere. The reaction was monitored by LCMS. The aqueous layer wasextracted with EtOAc. The resulting liquid was dried under vacuum. Thecrude product was used in the next step directly without furtherpurification. ESI-MS m/z: 374.20 [M+H]⁺.

Into a 250 mL round-bottom flask were added the compound from the step f(3.5 g, 9.35 mmol), CDI and THE (50 mL) at room temperature. Theresulting mixture was stirred for 2h at room temperature under nitrogenatmosphere. Then the resulting mixture was added to NH₃H₂O (400 mL)dropwise and stirred for 2h. The reaction was monitored by LCMS. Theaqueous layer was extracted with EtOAc, The residue was purified byreverse flash chromatography to afford the desired compound (1 g,28.65%) as white solid. ESI-MS m/z: 372.90 [M+H]⁺.

Intermediate 107

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. (95 mg, 91%) ESI-MS m/z:470.16 [M+H]⁺.

Intermediate 108

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. (476 mg, 86%) ESI-MS m/z:452.13 [M+H]⁺.

Intermediate 109

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. (382 mg, 68%) ESI-MS m/z:470/10 [M+H]⁺.

Intermediate 110

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, MeOH in DCM with NH₃) to afford thedesired product (305 mg, 59%) as a white solid. ESI-MS m/z: 434.15[M+H]⁺.

Intermediate 111

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material waspurified by prep-TLC (silica gel, MeOH in DCM with NH₃) to afford thedesired product (170 mg, 72%) as a white solid. ESI-MS m/z: 406.16[M+H]⁺.

Intermediate 112

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material wasconcentrated to dryness, redissolved in EtOAc, washed with NaHCO₃ (sat.aq.) 5×1 mL. The organic layer was dried over Na₂SO₄ filtered andconcentrated to afford the desired product (1.311 g, 80%) as a whitepowder. ESI-MS m/z: 436.23 [M+H]⁺.

Intermediate 113

The above compound was prepared in an analogous fashion to Intermediate47 with the corresponding aryl boronic acid. The crude material wasconcentrated to dryness, redissolved in EtOAc, washed with NaHCO₃ (sat.aq.) 5×1 mL. The organic layer was dried over Na₂SO₄ filtered andconcentrated to afford the desired product (589.7 mg, 82%) as a yellowpowder. ESI-MS m/z: 408.29 [M+H]⁺.

The following Table 8 contains examples that were prepared with thesimilar method to Example 1 Step c (PyBOP or HATU). The majority ofcompounds were purified by prep-HPLC (ACN/H₂O, 20-90%, 25 min), and somewere purified by automated column chromatography (silica gel). The arylacid and amine coupling partners were prepared according toIntermediates 1-113, or by analogous procedures with slightmodifications, and also prepared according to procedures found in U.S.patent application Ser. No. 16/930,622.

TABLE 8 Example Structure MS⁺ m/z 1067

585.20 1068

642.15 1069

636.20 1070

633.25 1071

667.20 1072

657.20 1073

639.20 1074

667.25 1075

659.40 1076

693.35 1077

683.40 1078

665.40 1079

694.45 1080

657.19 1081

673.20 1082

685.13 1083

685.13 1084

673.12 1085

666.17 1086

720.07 1087

684.19 1088

684.17 1089

702.16 1090

699.07 1091

655.12 1092

685.11 1093

673.10 1094

703.18 1095

654.03 1096

688.14 1097

694.81 1098

676.09 1099

670.12 1100

636.17 1101

654.09 1102

643.26 1103

673.26 1104

653.31 1105

625.1 1106

702.97 1107

648.34 1108

572.38 1109

598.38 1110

662.31 1111

688.32 1112

666.28 1113

707.08 1114

707.08 1115

677.12 1116

739.09 1117

649.18 1118

667.21 1119

652.15 1120

658.19 1121

699.01 1122

620.20 1123

624.20 1124

671.10 1125

637.08 1126

684.98 1127

720.94 1128

608.20 1129

703.20 1130

711.20 1131

629.20 1132

655.20 1133

636.29 1134

610.27 1135

650.26 1136

660.20 1137

660.20 1138

660.20 1139

620.24 1140

669.32 1141

639.18 1142

671.22 1143

616.23 1144

588.30 1145

622.31 1146

632.28 1147

640.30 1148

626.26 1149

671.16 1150

685.17 1151

641.23 1152

643.26 1153

611.25 1154

701.16 1155

638.19 1156

648.25 1157

656.18 1158

642.14 1159

657.14 1160

656.18 1161

666.14 1162

666.14 1163

638.19 1164

648.15 1165

656.18 1166

642.14 1167

608.18 1168

657.14 1169

701.09 1170

686.25 1171

642.22 1172

654.22 1173

696.19 1174

670.22 1175

700.26 1176

668.24 1177

638.23 1178

648.20 1179

642.18 1180

656.23 1181

627.12 1182

657.15 1183

659.19 1184

701.08 1185

721.04 1186

661.27 1187

627.23 1188

630.23 1189

683.06 1190

628.23 1191

638.23 1192

645.24 1193

617.25 1194

669.15 1195

623.38 1196

623.39 1197

674.27 1198

656.20 1199

656.20 1200

641.19 1201

680.14 1202

680.14 1203

680.16 1204

720.07 1205

720.19 1206

686.99 1207

704.97 1208

675.20 1209

657.24 1210

670.20 1211

641.18 1212

674.18 1213

659.21 1214

659.18 1215

641.20 1216

689.94 1217

673.20

Intermediate 114

To an oven-dried vial equipped with a stir bar was added6-bromo-2,2,3,3-tetrafluoro-2,3-dihydrobenzofuran (46 mg, 0.17 mmol),and 1,4-dioxane (0.85 mL). Then, Pd(dppf)Cl₂ (13 mg, 0.018 mmol),bis(pinacolato)diboron (65 mg, 0.26 mmol), and potassium acetate (33 mg,0.34 mmol) were added, and the reaction mixture was sparged with N₂ for5 minutes. The vial was then re-sealed and heated to 90° C. After 3hours, the reaction mixture was cooled to room temperature and filteredthrough a pad of silica gel using EtOAc to rinse. The filtrate wasconcentrated to afford crude boronic ester X as a dark brown oil (50 mg,assuming 100% yield) which was used without further purification.

Intermediate 115

The following intermediate was prepared in an analogous fashion toExample 890 to afford the desired product as light-yellow solid (1.63 g,62%) as a mixture of diastereomers. ESI-MS m/z: 572.28 [M+H]⁺.

The following Table 9 contains examples that were prepared with asimilar method to Example 891 using Example 890 and Intermediate 115. Ifnecessary to push conversion, more palladium and boronic acid wereadded. The majority of compounds were purified and the diastereomerswere separated by prep-HPLC (ACN/H₂O, 20-90%, 25 min). If reported as amixture of diastereomers, they were likely not separable by HPLC.

TABLE 9 Example Structure MS⁺ m/z 1218

726.11 1219

726.11 1220

712.18 1221

712.18 1222

712.08 1223

712.08 1224

694.06 1225

694.06 1226

710.16 1227

710.17 1228

636.23 1229

636.27 1230

682.20 1231

682.23 1232

632.27 1233

632.21 1234

666.14 1235

666.03 1236

710.20 1237

682.20 1238

682.30 1239

701.39 1240

602.20 1241

701.40 1242

666.23 1243

666.14 1244

666.19 1245

666.16 1246

650.29 1247

650.29 1248

630.79 1249

673.34 1250

673.29 1251

672.31 1252

649.15 1253

664.15 1254

664.15 1255

664.16 1256

664.19 1257

682.11 1258

682.20 1259

682.20 1260

682.20 1261

666.14 1262

682.20 1263

682.20 1264

682.20 1265

682.20 1266

682.20 1267

682.20

Example 1268

Example 1228 (15 mg, 0.024 mmol) was dissolved in EtOH (0.5 mL) and MeOH(0.5 mL) and palladium on carbon (10% wt., 0.8 mg, 0.00075 mmol) wasadded. The air in the headspace of the reaction vessel was replaced withH₂ from a balloon, and the mixture was allowed to stir under H₂ for 24hours. The reaction mixture was then filtered through a short pad ofcelite, and the filtrate concentrated to afford the product. (14 mg,93%). ESI-MS m/z: 638.29 [M+H]⁺.

Example 1269

In a vial, Example 890 (30 mg, 0.05 mmol) was dissolved in DMF (1 mL).Morpholine (44 mg, 0.5 mmol) and K₂CO₃ (69 mg, 0.5 mmol) were added andthe vial was sealed and heated to 100° C. for 24 h. The reaction wascooled to room temperature and diluted with H₂O (1 mL). The aqueouslayer was washed with EtOAc and the combined organic layer was driedover MgSO₄ and concentrated under reduced pressure. The crude materialwas purified by prep-HPLC (ACN/H₂O, 20-90%, 25 min) to give the titlecompound (5 mg, 16%). ESI-MS m/z: 605.27 [M+H]⁺.

Example 1270

The following example was prepared in an analogous fashion to Example1269. ESI-MS m/z: 639.20 [M+H]⁺.

Example 1271

The following example was prepared in an analogous fashion to Example1269. ESI-MS m/z: 639.20 [M+H]⁺.

The following Table 10 contains examples that were prepared with thesimilar method to Example 1 Step c (PyBOP or HATU) and Example 891 usingExample 890 and Intermediate 115. The majority of compounds werepurified by prep-HPLC (ACN/H₂O, 20-90%, 25 min), and some were purifiedby automated column chromatography (silica gel). The aryl acid and aminecoupling partners were prepared according to Intermediates 1-113, or byanalogous procedures with slight modifications, and also preparedaccording to procedures found in U.S. patent application Ser. No.16/930,622.

TABLE 10 Example Structure MS⁺ m/z 1272

704.04 1273

622.31 1274

622.29 1275

654.24 1276

604.32 1277

604.32 1278

640.29 1279

636.18 1280

636.23 1281

636.24 1282

604.23 1283

620.26 1284

666.16 1285

654.20 1286

636.18 1287

660.07 1288

644.15 1289

654.12 1290

600.29 1291

639.19 1292

635.22 1293

665.20 1294

682.30 1295

642.30 1296

636.35 1297

636.35 1298

654.35 1299

654.35 1300

654.35 1301

670.45 1302

624.35 1303

670.30 1304

670.35 1305

654.30 1306

621.10 1307

670.30 1308

621.30 1309

621.30 1310

648.32 1311

665.27 1312

648.31 1313

666.31 1314

693.19 1315

674.23 1316

665.26 1317

747.18 1318

737.03 1319

654.22 1320

628.20 1321

646.18 1322

638.24 1323

646.18 1324

647.15 1325

664.32 1326

646.30 1327

663.27 1328

646.31 1329

664.27 1330

682.27 1331

681.25 1332

664.27 1333

682.24 1334

664.28 1335

681.20

The following examples in Table 11 are prepared by using proceduressimilar to those described above:

Compound Structure P1

P2

P3

P4

P5

P6

P7

P8

P9

P10

P11

P12

P13

P14

P15

P16

P17

P18

P19

P20

P21

P22

P23

P24

P25

P26

P27

P28

P29

P30

P31

P32

P33

P34

P35

P36

P37

P38

P39

P40

P41

P42

P43

P44

P45

P46

P47

P48

P49

P50

P51

P52

P53

P54

P55

P56

P57

P58

P59

P60

P61

P62

P63

P64

P65

P66

P67

P68

P69

P70

P71

P72

P73

P74

P75

P76

P77

P78

P79

P80

P81

P82

P83

P84

P85

P86

P87

P88

P89

P90

P91

P92

P93

P94

P95

P96

P97

P98

P99

P100

P101

P102

P103

P104

P105

P105

P106

P107

P108

P109

P110

P111

P112

P113

P114

P115

P116

P117

P118

P119

P120

P121

P122

P123

P124

P125

P126

P127

P128

P129

P130

P131

P132

P133

P134

P135

P136

P137

P138

P139

P140

P141

P142

P143

P144

P145

P146

P147

P148P

P149

P150

P151

P152

P153

P154

P155

P156

P157

P158

P159

P160

P161

P162

P163

Assays Methods for RSV-A Assay

Hep-2 cells, (originally derived from tumors grown inirradiated-cortisonised weanling rats that had been injected withepidermoid carcinoma tissue from the larynx of a 56 year old male, butlater found to be indistinguishable from HeLa cells by PCR DNAanalysis), were used for the culturing of genotype A, “Long” strain RSV.Flasks were inoculated with RSV and viral stocks were collected oncecytopathic effect (CPE) was greater than 90%. Viral stocks in 25%sucrose media were snap frozen using liquid nitrogen to increase viralstability. Viral stock titers were quantified by tissue cultureinfectious dose 50% (TCID₅₀) using 8,000 cells per well and 3-fold viraldilutions across a 96-well plate, cultured for 4 days. Viral stocktiters were also quantified by a plaque forming unit assay, as describedelsewhere.

Following extensive parameter testing, the final assay is run asfollows: Hep-2 cells are seeded into the inner 60 wells of a 96-wellplate at 8,000 cells per well in a volume of 50 μL using Growth Media(DMEM without phenol red, 1% L-Glut, 1% Penn/Strep, 1% nonessentialamino acids, 10% heat-inactivated FBS). 2-fold serial dilutions ofcontrol and test compounds are added to the wells in duplicate in atotal volume of 25 μL. Viral stock is then added to the wells at amultiplicity of infection (MOI) of 0.1 in a volume of 25 μL, bringingthe total volume of each well to 100 μL. The MOI is calculated using thePFU/mL, or TCID₅₀ if unavailable. Each 96-well plate has a controlcolumn of 6 wells with cells and virus but no compound (negativecontrol, max CPE), a column with cells but no compound or virus(positive control, minimum CPE), and a column with no cells or virus orcompound (background plate/reagent control). The control wells withcells but no virus are given an additional 25 μL of growth mediacontaining an equal quantity of sucrose as those wells receiving theviral stock in order to keep consistent in media and volume conditions.The outer wells of the plate are filled with 125 μL of moat media (DMEM,1% Penn/Strep) to act as a thermal and evaporative moat around the testwells. Following a 5-day incubation period, the plates are read usingATPlite (50 uL added per well), which quantifies the amount of ATP (ameasure of cell health) present in each well. Assay plates are readusing the Envision luminometer. These data are used to calculate theEC₅₀ each compound (Table 12). EC₅₀ ranges are as follows: A<0.2 μM;B>0.2 μM.

TABLE 12 Summary of Activities for RSV-A Human Human RSV-A RSV-ACompound (“Long”) Compound (“Long”) 3 A 4 A 5 B 6 A 7 A 8 A 9 A 10 A 11A 12 A 13 A 14 A 15 A 16 A 17 A 18 A 19 A 20 A 21 A 22 A 23 A 24 A 25 A26 A 27 A 28 A 29 A 30 A 31 B 32 A 33 B 34 A 35 A 36 A 37 A 38 B 39 B 40A 41 A 42 A 43 A 44 A 45 A 46 A 47 A 48 A 49 A 50 A 51 A 52 A 53 A 54 A55 A 56 B 57 A 58 A 59 A 60 A 61 A 62 A 63 A 64 A 65 A 66 B 67 A 68 A 69A 70 A 71 A 72 A 73 A 74 A 75 A 76 B 77 A 78 A 79 B 80 A 81 A 82 A 83 A84 A 85 A 86 A 87 A 88 A 89 A 90 A 91 A 92 A 93 A 94 A 95 A 96 A 97 A 98A 99 A 100 A 101 A 102 A 103 A 104 A 105 A 106 B 107 B 108 A 109 B 110 B111 B 112 A 113 A 114 A 115 A 116 A 117 A 118 A 119 A 120 A 121 A 122 A123 A 124 A 125 B 126 B 127 B 128 A 129 A 130 A 131 A 132 A 133 A 134 A135 A 136 A 137 A 138 A 139 A 140 B 141 A 142 A 143 A 144 B 145 B 146 B147 A 148 A 149 A 150 B 151 B 152 A 153 A 154 A 155 A 156 A 157 A 158 A159 A 160 A 161 A 162 B 163 A 164 A 165 A 166 A 167 A 168 A 169 A 170 A171 A 172 B 173 A 174 A 175 B 176 B 177 A 178 A 179 A 180 B 181 B 182 A183 A 184 A 185 B 186 B 187 A 188 A 189 B 190 B 191 A 192 B 193 B 194 A195 A 196 B 197 A 198 B 199 A 200 A 201 B 202 A 203 A 204 A 205 B 206 A207 B 208 B 209 B 210 A 211 A 212 B 213 B 214 B 215 A 216 A 217 A 218 A219 A 220 A 221 A 222 A 223 A 224 A 225 A 226 A 227 A 228 A 229 A 232 A233 A 234 A 235 A 236 A 237 A 238 B 239 A 240 A 241 A 242 A 243 B 244 B245 A 246 B 247 B 248 B 249 B 250 B 251 B 252 A 253 A 254 A 255 A 256 A257 A 258 A 259 A 260 A 261 A 262 B 263 A 264 B 265 A 266 B 267 A 268 A269 B 270 A 271 B 272 A 273 B 274 A 275 A 276 A 277 B 278 A 279 B 280 A281 B 282 B 283 A 284 A 285 A 286 A 287 A 288 A 289 B 290 A 291 B 292 A293 B 294 A 295 A 296 B 297 A 298 B 299 A 300 A 301 A 302 A 303 A 304 A305 A 306 A 307 A 308 A 309 A 310 B 311 B 312 A 313 A 314 A 315 A 316 A317 A 318 A 319 A 320 A 321 A 322 A 323 A 324 A 325 A 326 A 327 A 328 A329 A 330 B 331 A 332 A 333 B 334 B 335 B 336 B 337 B 338 B 339 B 340 A341 B 342 B 343 B 344 A 345 B 346 B 347 B 348 A 349 B 350 B 351 B 352 B353 B 355 B 356 B 357 B 358 B 359 B 360 B 361 B 362 B 363 B 364 B 365 B366 A 367 B 368 B 369 A 370 A 371 B 372 B 373 B 374 B 375 B 376 B 377 B378 B 379 B 380 A 381 B 382 B 383 B 384 B 385 B 386 B 387 B 388 B 389 B390 B 391 B 392 B 393 B 394 B 395 B 396 B 397 B 398 B 399 B 400 B 401 B402 B 403 B 404 B 405 B 406 B 407 B 408 B 409 B 410 B 411 B 412 B 413 B414 B 415 B 416 B 417 B 418 A 419 B 420 B 421 B 422 B 423 B 424 B 425 B426 B 427 B 428 B 429 B 430 B 431 B 432 B 433 B 434 B 435 B 436 B 437 B438 B 439 B 440 A 441 A 442 B 443 B 444 A 445 B 446 B 447 B 448 B 449 B450 B 451 B 452 A 453 B 454 A 455 B 456 B 457 B 458 A 459 B 460 A 461 A462 A 463 A 464 A 482 B 483 B 484 B 485 B 486 A 487 A 488 A 489 B 490 A491 A 492 A 493 A 494 A 495 A 496 A 497 A 498 A 499 A 500 A 501 A 502 A503 A 504 A 505 A 506 A 507 A 508 A 509 A 510 A 511 A 512 A 513 A 514 A515 A 516 A 517 A 518 A 519 A 520 A 521 A 522 A 523 A 524 A 525 A 526 A527 A 528 A 529 A 530 A 531 A 532 A 533 A 534 A 535 A 536 A 537 A 538 A539 A 540 A 541 A 542 A 543 A 544 A 545 A 546 A 547 A 548 A 549 A 550 A551 A 552 A 553 A 554 A 555 A 556 A 557 A 558 A 559 A 560 A 561 A 562 A563 A 564 A 565 A 566 A 567 A 568 A 569 A 570 A 571 A 572 A 573 A 574 A575 A 576 A 577 A 578 A 579 A 580 A 581 A 582 A 583 A 584 A 585 A 586 A587 A 588 A 589 A 590 A 591 A 592 A 593 A 594 A 595 A 596 A 597 A 598 A599 A 600 A 601 A 602 A 603 A 604 A 605 A 606 A 607 A 608 A 609 A 610 A611 A 612 A 613 A 614 A 615 A 616 A 617 A 618 A 619 A 620 A 621 A 622 A623 A 624 A 625 A 626 A 627 A 628 B 633 A 634 A 635 B 636 A 637 A 638 A639 A 640 A 641 A 642 A 643 A 644 B 645 A 646 A 647 A 648 A 649 A 650 A651 A 652 A 653 A 654 A 655 A 656 A 657 A 658 A 659 A 660 A 661 A 662 A663 A 664 A 665 A 666 A 667 A 668 A 669 A 670 A 671 A 672 A 673 A 674 A675 A 683 A 684 A 685 A 686 A 687 A 688 A 691 A 692 B 693 A 694 A 695 A696 A 697 A 698 A 699 A 700 A 701 A 702 A 703 A 704 A 705 A 706 A 707 A708 A 709 A 710 A 711 A 712 A 713 A 714 A 715 A 716 A 717 A 718 A 719 A720 A 721 A 722 A 723 A 724 A 725 A 726 A 727 A 728 A 729 A 730 A 731 A732 A 733 A 734 A 735 A 736 A 737 A 738 A 739 A 740 A 741 A 742 A 743 A744 A 745 A 746 A 747 A 748 A 749 A 750 A 751 A 752 A 753 A 754 A 755 A756 A 757 A 758 A 759 A 760 A 761 A 762 A 763 A 764 A 765 A 766 A 767 A768 A 773 A 774 A 775 A 776 A 777 A 778 A 779 A 780 A 781 A 782 A 783 A784 A 785 A 786 A 787 A 788 A 789 A 790 A 791 A 792 A 793 A 794 A 795 A796 A 797 A 798 A 799 A 800 A 801 A 802 A 805 B 806 B 807 B 808 A 809 B810 B 811 A 812 B 813 B 814 B 815 A 816 A 817 A 818 A 819 A 820 A 821 A822 A 823 A 824 A 825 A 826 A 827 A 828 A 829 A 830 A 831 A 832 A 833 A834 A 835 A 836 A 837 A 838 A 839 A 840 A 841 A 842 A 843 A 844 A 845 A846 A 847 A 848 A 849 A 850 A 851 A 852 A 853 A 854 B 855 A 856 A 857 A858 A 859 A 860 A 861 A 862 A 871 A 872 A 873 A 874 B 875 B 876 B 877 A878 A 879 B 880 B 881 A 882 B 883 A 884 A 885 A 886 A 887 A 888 A 889 A890 B 891 A 892 B 893 A 894 B 895 B 896 A 897 B 898 A 899 B 900 A 901 A902 B 903 A 904 B 905 B 906 A 907 B 908 A 909 A 910 B 911 A 912 A 913 B914 A 915 A 916 A 917 A 918 A 919 B 920 A 921 B 922 A 923 B 924 A 925 A926 A 927 A 928 A 929 A 930 A 931 A 932 A 933 A 934 A 935 A 936 A 937 B938 B 939 A 940 A 941 B 942 B 943 A 944 A 945 A 946 A 947 A 948 A 949 A950 A 951 A 952 A 953 A 954 A 955 A 956 A 961 A 965 B 966 B 967 B 968 B969 B 970 B 971 B 972 A 973 B 974 A 975 A 976 A 977 A 978 A 979 A 980 A981 B 982 A 983 A 984 B 985 A 986 A 987 A 988 A 989 A 990 A 991 A 992 A993 A 994 A 995 B 996 A 997 A 998 A 999 A 1000 A 1001 A 1002 B 1003 A1004 B 1005 A 1006 B 1007 A 1008 B 1009 A 1010 B 1011 A 1012 B 1013 A1014 A 1015 A 1016 B 1017 A 1018 B 1019 A 1020 B 1021 A 1022 B 1023 A1024 B 1025 A 1026 B 1027 A 1028 B 1029 A 1030 A 1031 A 1032 B 1033 A1034 B 1035 A 1036 B 1037 A 1038 B 1039 A 1040 A 1041 A 1042 A 1043 A1044 A 1045 A 1046 B 1047 A 1048 B 1049 A 1050 A 1051 A 1052 A 1053 B1054 A 1055 B 1056 A 1057 B 1058 A 1059 B 1060 A 1061 B 1062 A 1063 B1064 A 1065 A 1066 B 1067 A 1068 A 1069 A 1071 A 1072 A 1073 A 1074 A1075 A 1076 A 1077 A 1078 A 1079 A 1080 A 1081 A 1082 A 1083 A 1084 A1085 A 1086 A 1087 A 1088 A 1089 A 1090 A 1091 A 1092 A 1093 A 1094 A1095 A 1096 A 1097 A 1098 A 1099 A 1100 A 1101 A 1102 A 1103 A 1104 B1105 A 1106 A 1107 A 1108 A 1109 A 1110 A 1111 A 1112 A 1113 A 1114 A1115 A 1116 A 1117 A 1118 A 1119 A 1120 A 1121 A 1122 A 1123 A 1124 A1125 A 1126 A 1127 A 1128 A 1129 A 1130 A 1131 A 1132 A 1133 A 1134 A1135 A 1136 A 1137 A 1138 A 1139 A 1140 A 1141 A 1142 A 1143 A 1144 A1145 A 1146 A 1147 A 1148 A 1149 A 1150 A 1151 A 1152 A 1153 A 1154 A1155 A 1156 A 1157 A 1158 A 1159 A 1160 A 1161 A 1162 A 1163 A 1164 A1165 A 1166 A 1167 A 1168 A 1169 A 1170 A 1171 A 1172 A 1173 A 1174 A1175 A 1176 A 1177 A 1178 A 1179 A 1180 A 1181 A 1182 A 1183 A 1184 A1185 A 1186 A 1187 A 1188 A 1189 A 1190 A 1191 A 1192 A 1193 A 1194 A1195 A 1196 A 1197 A 1198 A 1199 A 1200 A 1201 A 1202 A 1203 A 1204 A1205 A 1206 A 1207 A 1208 A 1209 A 1210 A 1211 A 1212 A 1213 A 1214 A1215 A 1216 A 1217 B 1218 A 1219 A 1220 A 1221 A 1222 A 1223 A 1224 A1225 A 1226 A 1227 A 1228 A 1229 B 1230 B 1231 A 1232 B 1233 A 1234 B1235 A 1236 B 1237 B 1238 A 1239 B 1240 B 1241 B 1242 A 1243 A 1244 A1245 A 1246 A 1247 A 1248 A 1249 B 1250 A 1251 B 1252 A 1253 A 1254 A1255 A 1256 A 1257 A 1258 A 1259 A 1260 A 1261 A 1262 A 1263 A 1264 A1265 A 1266 A 1267 A 1268 A 1269 B 1270 A 1271 A 1272 A 1273 A 1274 A1275 A 1276 A 1277 A 1278 A 1279 A 1280 A 1281 A 1282 A 1283 A 1284 A1285 A 1286 A 1287 A 1288 A 1289 A 1290 A 1291 A 1292 B 1293 A

Methods for HMPV Antiviral Assay Method-A:

HMPV antiviral activity was evaluated using a recombinant version ofHMPV CAN97-83 engineered to contain the coding sequence for enhancedgreen fluorescence protein (eGFP) in the 3′ end of the virus genome(MPV-GFP1, ViraTree). Vero E6 cells (ATCC #CCL-7) were seeded at adensity of 12,000 cells/100 μL/well into 96-well cell plates one dayprior to the assay. On the day of screening, the cell culture medium wasaspirated from the wells and cells were washed twice with serum-freeEagle's Modified Essential Medium (EMEM, ATCC #) containing 1%penicillin-streptomycin (Invitrogen) (SF-EMEM). Cell washes wereperformed by dispensing 100 μL SF-EMEM per well and immediatelyaspirating the wash medium from the well. Following the second washstep, serum-free OptiMEM (Invitrogen, Cat No.) (SF-OptiMEM) containing0.5 μg/mL TPCK-Trypsin (VENDOR) and 1% penicillin-streptomycin was addedto the cells at 50 μL/well. Compounds were added into the 96-well platesusing a JANUS automated liquid handling system (VENDOR). Compounds wereinitially diluted 1:50 into an intermediate 96-well plate containingSF-OptiMEM prior to transfer to the assay plate (25 μL/well). Each ofthe test compounds were tested in duplicate wells at finalconcentrations starting from 8 μM or 2 μM using ½ stepwise dilutions fora total of 8 points. Virus infection was performed by preparing aworking stock of MPV-GFP1 at a multiplicity of infection (MOI) equal to0.05/25 μL and aliquoting 25 μL of virus inoculum to the compound andpositive control wells. SF-OptiMEM was added (25 μL/well) to theappropriate wells to serve as a virus-free negative control for theassay. The final DMSO concentration of all wells is 0.5%. Plates wereincubated at 32° C., 5% CO₂ for 5 days.

After 5 days incubation, eGFP fluorescence intensity was measured at (X)nM wavelength using a Spectramax i3× plate reader (VENDOR). Percentviral inhibition was calculated using the following equation:

y=[100−(X _(Q) /X _(P))]×100

Where X_(Q) is the fluorescence intensity measured in a well containingrecombinant MPV-GFP1-infected, compound-treated cells and X_(P) is theaverage fluorescence intensity measured in the wells containinguntreated cells infected with recombinant virus. EC₅₀ values were thencalculated by non-linear regression using a four parameter curvelogistic equation. The curve fit model employed was XLFit Dose ResponseOne Site Model 200:

y=(A+(B/(1+((x/C){circumflex over ( )}D))))

Where A is the minimum y value, B is the maximum y value, C is the logEC₅₀ value, and D is the slope factor. These data are used to calculatethe EC₅₀ each compound (Table 13). EC₅₀ ranges are as follows: A<0.5 μM;B>0.5 μM.

TABLE 13 Summary of Activities for HMPV HMPV HMPV Compound EC₅₀ CompoundEC₅₀ 3 B 4 A 6 A 7 A 8 A 9 A 10 A 11 B 12 A 13 A 14 A 16 A 17 A 19 A 20A 22 A 23 A 24 A 25 A 26 A 27 A 28 A 29 A 30 A 32 A 34 B 40 A 41 A 42 A43 A 44 A 45 A 47 A 48 A 49 B 50 B 51 A 52 A 53 A 54 B 55 A 57 B 58 A 59A 60 A 62 B 63 B 64 B 65 B 68 A 70 A 72 A 73 A 74 A 75 B 77 A 78 B 80 A81 B 82 A 83 A 84 A 85 A 86 A 91 B 92 A 94 B 95 B 96 A 97 B 100 B 102 A103 A 105 B 108 B 112 B 113 A 114 A 120 B 121 A 124 A 128 A 129 B 132 B134 B 135 B 136 B 137 B 138 B 139 B 141 B 142 B 143 A 147 A 148 A 149 B152 B 153 B 154 B 155 B 156 B 157 B 158 B 160 B 161 B 165 B 166 B 167 B168 A 169 B 170 A 174 B 177 A 178 B 179 B 182 A 183 B 184 B 199 B 200 B210 A 217 A 218 A 219 A 220 B 221 B 222 B 223 B 224 B 226 B 234 A 235 A263 A 265 A 267 A 268 B 270 A 280 A 300 A 301 A 305 A 306 A 307 A 308 A309 B 314 A 315 A 321 A 322 B 323 B 324 A 327 B 329 A 441 B

Method-B:

In vitro HMPV antiviral activity was evaluated using the clinicalisolate TN/1501/A1 and LLC-MK2 cells (ATCC #CCL-7), an immortalizedkidney epithelial cell line from Macaca mulatta.

Compounds were resuspended in dimethyl sulfoxide (DMSO) at 10 mM andadded to a 384-well source plate. The compounds were diluted andtransferred onto 384-well assay plates using the Echo-650 automatedliquid handling system (Beckman Coulter, Ind.). The test compounds wereassessed in duplicate at a top concentration of 10 μM followed by 3-foldserial dilutions to give a total of 10 concentration points. DMSOcontrol wells were also included on the assay plates and were eitherinfected or not, acting as positive and negative controls.

TN/1501/A1 virus infections were performed in-suspension with LLC-MK2cells. The cells were washed twice with PBS and removed from thecell-culture flask with 0.25% trypsin-EDTA (Thermo Fisher Scientific,MA). The trypsin-EDTA was inactivated by resuspending in 2% fetal bovineserum (FBS) and OptiMEM (ThermoFisher Scientific, MA) containing 1%penicillin-streptomycin. Cells were pelleted by centrifuging for 5minutes at 800 rpm, the supernatant was removed, and cells weresuspended in PBS plus 100 μg/mL CaCl₂). This step was performed twice.Cells were then resuspended in serum-free (SF)-OptiMEM containing 4μg/mL TPCK-Trypsin (Sigma Aldrich, MO), 1% penicillin-streptomycin(ThermoFisher Scientific, MA) and 100 μg/mL CaCl₂). Cells were countedand seeded at a density of 5,000 cells/well, 12.5 μL/well.

Virus infections were done at a multiplicity of infection (MOI) of 0.014with 12.5 μL added per well. SF-OptiMEM, 12.5 μL/well, was added to theappropriate wells to serve as a virus-free negative control for theassay. The final concentration of TPCK-trypsin was 2 μg/mL. Plates wereincubated at 37° C., 5% CO₂ for 6 days.

After 6 days incubation, 12.5 μL of ATP-Lite (PerkinElmer, MA) was addedto each well and the raw luminescence values were determined using theEnvision 2104 (Perkin Elmer, MA). The average of the raw luminescencevalues for the cells and virus only positive control wells wassubtracted from all conditions tested and the percent cell health wasdetermined by dividing these values by the average of the cells onlynegative control wells. EC₅₀ values were then calculated by non-linearregression using a four-parameter curve logistic equation. The curve fitmodel employed was XLFit Dose Response One Site Model 200:

y=(A+(B/(1+((x/C){circumflex over ( )}D)))),

where A is the minimum y value, B is the maximum y value, C is the logEC₅₀ value, and D is the slope factor. These data are used to calculatethe EC₅₀ each compound (Table 14). EC₅₀ ranges are as follows: A<0.5 μM;B>0.5 μM.

TABLE 14 Summary of Activities for hMPV WT1501 HMPV HMPV Compound EC₅₀Compound EC₅₀ 486 B 487 B 488 B 489 B 491 A 493 A 494 A 495 A 496 A 497A 498 A 499 A 500 A 501 B 502 A 503 A 504 A 505 A 506 A 507 A 508 A 509A 510 A 511 B 512 B 513 A 514 A 515 A 516 B 517 A 518 A 519 B 520 A 521A 522 A 523 A 524 A 525 A 526 A 527 A 528 A 529 A 530 A 531 A 532 A 533A 534 A 535 A 536 A 537 A 538 A 539 A 540 A 541 A 542 A 543 A 544 A 545A 546 A 547 A 548 A 549 A 550 A 551 A 552 A 553 A 554 A 555 A 556 A 557A 558 A 559 A 560 A 561 A 562 A 563 A 564 A 565 A 566 A 567 A 568 A 569A 570 A 571 A 572 A 573 A 574 A 575 A 576 B 577 B 578 B 579 B 580 B 581B 582 B 583 B 585 B 586 B 587 B 588 B 589 B 590 B 591 A 592 A 593 A 594A 595 A 596 A 597 B 598 A 599 A 600 A 601 A 602 A 603 B 604 A 605 A 606A 607 A 608 A 609 A 610 A 611 A 612 A 613 A 614 B 615 A 616 A 617 A 618A 619 A 620 A 621 A 622 A 623 B 624 B 625 B 626 B 627 B 628 B 629 A 630A 631 A 632 A 633 A 636 A 637 A 638 A 639 A 640 A 641 A 642 B 643 A 644B 645 B 646 B 647 B 648 B 649 B 650 A 651 A 652 A 653 B 654 A 655 A 656A 657 A 658 A 659 A 660 A 661 A 662 A 663 A 664 A 665 A 666 A 667 A 668A 669 B 670 B 671 B 672 B 673 B 674 B 675 B 676 A 677 B 678 A 679 B 680B 681 B 682 B 683 A 684 A 685 A 686 A 687 A 688 A 689 A 690 A 691 B 693B 694 B 695 B 696 B 697 A 698 B 699 B 700 B 702 B 703 A 704 A 705 A 706A 707 A 708 A 709 B 710 A 711 A 712 A 713 A 714 A 715 A 716 A 717 A 718B 719 A 720 A 721 B 722 A 723 A 724 A 725 A 726 A 727 A 728 A 729 A 730A 731 A 732 A 733 A 734 A 735 A 736 A 737 A 738 A 739 A 740 A 741 A 742A 743 A 744 A 745 A 746 A 747 A 748 A 749 A 750 A 751 A 752 A 753 A 754A 755 A 756 A 757 A 758 A 759 A 760 A 761 A 762 A 763 A 764 A 765 A 766A 767 A 768 A 769 A 770 A 771 A 772 A 773 A 774 A 775 A 776 A 777 A 778A 779 A 780 A 781 A 782 A 783 A 784 A 785 A 786 A 787 A 788 A 789 A 790A 791 A 792 A 793 A 794 A 795 B 796 A 797 A 798 A 799 A 800 A 801 A 802A 803 A 804 A 811 A 815 A 816 A 817 A 818 A 819 A 820 A 821 B 822 A 823A 824 A 825 A 826 A 827 A 828 A 829 B 830 A 831 A 832 B 833 B 834 A 835A 836 A 837 A 838 A 839 A 840 A 841 A 842 A 843 A 844 A 845 A 846 A 847A 848 A 849 A 850 B 851 A 852 B 853 A 854 B 855 A 856 A 857 A 858 A 859A 860 A 861 A 862 A 863 B 864 B 865 B 866 B 867 B 868 B 869 A 870 B 871A 872 A 873 A 874 B 876 B 877 A 878 B 883 B 884 B 885 A 886 A 887 B 888B 889 B 891 B 893 A 896 B 898 B 899 B 901 A 903 B 908 A 909 B 911 A 912A 913 B 914 B 915 A 916 A 917 B 918 B 919 B 920 A 921 B 922 A 923 B 924B 925 B 926 B 927 B 928 B 929 A 930 A 931 A 932 A 933 B 934 B 935 B 936A 937 B 938 B 939 B 940 B 941 B 942 B 943 B 944 B 945 B 946 A 947 B 948B 949 B 950 B 951 A 952 A 953 B 954 B 955 B 956 A 957 A 958 A 959 B 960A 961 A 962 A 963 A 964 A 965 B 966 B 967 B 968 B 969 B 970 B 971 B 972A 973 B 974 A 975 A 976 B 977 B 978 B 979 A 980 A 981 B 982 B 983 B 984B 985 A 986 A 987 A 988 A 989 A 990 A 991 A 992 A 993 A 994 A 995 B 996A 997 A 999 A 1000 B 1001 A 1002 B 1003 A 1004 B 1005 A 1006 B 1007 A1008 B 1009 A 1010 B 1011 A 1012 B 1013 A 1014 A 1015 A 1016 B 1017 A1018 B 1019 A 1020 B 1021 A 1022 B 1023 A 1024 B 1025 A 1026 B 1027 A1028 B 1029 A 1030 B 1031 A 1032 B 1033 A 1034 B 1035 A 1036 B 1037 A1038 B 1039 A 1040 B 1041 A 1042 A 1043 A 1044 A 1045 A 1046 B 1047 A1049 A 1050 A 1051 B 1052 A 1053 B 1054 B 1055 B 1056 A 1057 B 1058 A1059 B 1060 A 1061 B 1062 A 1063 B 1064 A 1065 A 1066 B 1067 B 1068 A1069 B 1072 A 1074 A 1080 A 1081 A 1082 A 1084 A 1085 A 1087 A 1088 A1089 A 1090 B 1091 A 1092 A 1093 A 1094 A 1095 A 1096 A 1097 A 1098 A1099 A 1100 A 1101 A 1102 A 1103 A 1104 B 1105 B 1106 A 1107 A 1108 A1109 A 1110 A 1111 A 1112 A 1113 A 1114 A 1115 A 1116 A 1117 A 1118 A1119 A 1120 A 1121 A 1122 A 1123 A 1124 A 1125 A 1126 A 1127 A 1128 B1133 B 1134 B 1135 A 1136 A 1137 A 1138 A 1139 A 1140 A 1141 A 1142 A1143 B 1144 B 1145 A 1146 A 1147 A 1148 A 1149 A 1150 A 1151 A 1152 A1153 A 1154 A 1155 A 1156 A 1157 A 1158 A 1159 A 1160 A 1161 A 1162 A1163 A 1164 A 1165 A 1166 A 1167 A 1168 A 1169 A 1170 B 1177 A 1178 A1179 A 1180 A 1181 A 1182 A 1183 A 1184 A 1185 A 1186 A 1187 A 1188 A1189 A 1190 A 1191 A 1192 B 1193 B 1194 B 1195 A 1196 B 1218 B 1219 B1220 B 1221 A 1222 B 1223 A 1224 B 1225 A 1226 B 1227 A 1228 B 1229 B1230 B 1231 A 1232 B 1233 B 1234 B 1235 A 1248 A 1249 B 1250 B 1251 B1269 B

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by one of Formulae (XXIX-1)˜(XXIX-3):

or a pharmaceutically acceptable salt thereof, wherein R₄₆ issubstituted methyl or optionally substituted cyclopropyl; R₄₇ ishydrogen, Cl, F, or optionally substituted methoxyl; R₄₁, R₄₂, R₄₃, R₄₄,or R₄₅ is each independently selected from hydrogen, halogen, optionallysubstituted methyl, optionally substituted methoxy or —CN;alternatively, R₄₂ and R₄₃, or R₄₁ and R₄₂ are taken together with thecarbon atoms to which they are attached to form a fused 4- to 7-memberedcarbocyclic ring or heterocyclic ring.
 2. The compound of claim 1,wherein R₄₆ cyclopropyl and R₄₇ is methoxy.
 3. The compound of claim 1,which is represented by Formula (XIX-11) or Formula (XIX-12),

or a pharmaceutically acceptable salt thereof, wherein: R₁₄ isoptionally substituted cyclopropyl; each R₂₁ is independently —F, —Cl,—CN, —CF₃, —CH₂F or —CHF₂; and R₂₂ is Cl, F, or optionally substitutedmethoxy.
 4. The compound of claim 3, wherein R₂₂ is methoxy.
 5. Thecompound of claim 1, represented by Formula (XXIII-9) or Formula(XXIII-10),

or a pharmaceutically acceptable salt thereof, wherein: R₄ is CF₃ orcyclopropyl; R₁₄ is optionally substituted cyclopropyl; each R₂₁ isindependently —F, —Cl, —CN, —CF₃, —CH₂F or —CHF₂; R₂₄ is hydrogen, Cl,F, or optionally substituted methoxy; and W is methyl.
 6. A compoundselected from the compounds set forth below, or a pharmaceuticallyacceptable salt thereof: Example Structure   3

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P163


7. A pharmaceutical composition comprising a compound according to claim1, or a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier, diluent or excipient.
 8. A method of treating orpreventing an RSV infection in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of acompound or a combination of compounds of claim
 1. 9. The method ofclaim 8, further comprising the step of administering to the subject anadditional anti-RSV agent.
 10. The method of claim 8, further comprisingadministering to the subject a steroid anti-inflammatory compound.
 11. Amethod of treating an RSV infection and influenza in a subject in needthereof, comprising administering to the subject a therapeuticallyeffective amount of a compound of claim 1 and a therapeuticallyeffective amount of an anti-influenza agent.
 12. The method of claim 9,wherein the compound and the additional anti-RSV agent areco-formulated.
 13. The method of claim 9, wherein the compound and theadditional anti-RSV agent are co-administered.
 14. The method of claim9, wherein administering the compound allows for administering of theadditional anti-RSV agent at a lower dose or frequency as compared tothe administering of the additional anti-RSV agent alone that isrequired to achieve similar results in prophylactically treating an RSVinfection in a subject in need thereof.
 15. A method of treating orpreventing an HMPV infection in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of acompound or a combination of compounds of claim
 1. 16. The method ofclaim 15, further comprising the step of administering to the subject anadditional anti-HMPV agent.
 17. The method of claim 16, wherein thecompound and the additional anti-HMPV agent are co-formulated.
 18. Themethod of claim 16, wherein the compound and the additional anti-HMPVagent are co-administered.